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HK1254830A1 - 工程化植酸酶及其使用方法 - Google Patents

工程化植酸酶及其使用方法 Download PDF

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HK1254830A1
HK1254830A1 HK18113922.6A HK18113922A HK1254830A1 HK 1254830 A1 HK1254830 A1 HK 1254830A1 HK 18113922 A HK18113922 A HK 18113922A HK 1254830 A1 HK1254830 A1 HK 1254830A1
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R‧M‧拉布
G‧拉扎尔
B‧沈
G‧拉扎爾
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谷万达公司
谷萬達公司
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Description

工程化植酸酶及其使用方法
相关申请的交叉引证
本申请要求于2015年9月18日提交的美国临时申请第62/220,688号的权益,如同所充分阐述的,其通过引证结合至本文。
与本申请一起电子提交的序列表以“序列表”为标题,创建于2016年9月18日,并且具有399,733字节的文件大小,如同所充分阐述的,该序列表通过引证结合至本文。
技术领域
本公开涉及工程化植酸酶分子(其具有改善的热稳定性,改善的或降低的胃稳定性),编码其的核酸,其制备方法,以及使用其在工业加工或动物饲料中的方法。
本公开涉及表达具有改善的热稳定性的植酸酶的转基因植物,编码该植酸酶的核酸,以及加工该转基因植物与组织,和生产和利用动物饲料的方法。本公开还涉及含有植酸酶的饲料添加剂、谷物和纤维加工添加剂。
本公开涉及工程化大肠杆菌来源的植酸酶的形式,其已经被改性以改善它们作为单胃和反刍动物饲料组分的性能。这些改性的植酸酶可以直接在饲料组分,如玉米粒中表达,并且掺入动物饲料中,例如掺入单胃动物的饲料(mash)或颗粒饲料中,或者掺入反刍动物的青贮饲料或谷物中。含有表达植酸酶的这些植物的日粮支持有效的动物生长,且相比没有工程化植酸酶时所必需的磷酸盐更少的磷酸盐。
背景技术
植酸酶是一类从植酸中水解磷酸盐以产生游离的磷酸盐和肌醇的酸性磷酸酶。植酸(肌醇六磷酸),或其去质子化形式,植酸盐在许多动物饲料组分,如谷物和豆类中很常见,并且可以代表这些饲料中总磷酸盐含量的显著部分。然而,许多家畜不能有效消化植酸,因此不能吸收磷酸盐。
因此,必须将其它形式的磷酸盐,如岩石磷酸盐或磷酸钙添加到动物饲料中以提供这种关键营养素。此外,植酸作为饮食中的抗营养元素,与蛋白质和螯合矿物质,如铁、钙和镁结合,阻止了这些物质的吸收。由于未消化的植酸和过量的无机磷酸盐可以排泄在粪便中,它们可以作为农业径流(agricultural run-off)中磷酸盐污染的重要来源。植酸酶通常用于工业加工和动物生产。在动物饲料中添加植酸酶可以降低添加无机磷酸盐的需求,增加动物对磷酸盐、蛋白质和矿物质的吸收,并减少农业径流中的磷酸盐污染。当这些效应结合起来时,可以显著提高动物生长的效率以及从它们所消耗的饲料中获得的总营养。
在工业过程中,特别是发酵过程中,植酸酶常用于水解植酸盐,使矿物质和其他营养物质释放进入发酵,以及通过需要辅因子(被植酸盐螯合的(sequestered))的酶来增强淀粉降解(E.Khullar,J.K.Shetty,M.E.Tumbleson,V.Singh,“Use of Phytases inEthanol Production from E-Mill Corn Processing,”Cereal Chem.,88(3):223-227,2011,如同所充分阐述的,其通过引证结合至本文)。它在工业上也用于减少可能与发酵或相关过程中经常发生的和植酸盐或磷酸盐积聚(有时称为“啤酒石”)有关的结垢。在动物生产和营养方面,对于营养学上可行的由植酸盐制备磷的一个策略是向动物饲料中添加植酸酶(Jongbloed和Lenis,1998;Onyango等,2005,如同所充分阐述的,两者均通过引证结合至本文)。已经表明,在家禽和猪的日粮中使用植酸酶提高了性能和磷利用率(Baker,2002;Nyannor等,2007和2009,如同所充分阐述的,两者都通过引证结合至本文)。目前市场上有多种植酸酶产品用于此用途,包括NatuphosTM(BASF),来源于黑曲霉的植酸酶,RonozymeTM(DSM),来源于隔孢伏革菌(Peniophora lycii)的植酸酶,以及来源于大肠杆菌的Quantum和Quantum Blue(AB Vista)植酸酶。在动物饲料中使用植酸酶可以减少添加到动物饲料中的无机磷的量,并且据报导,粪便磷降低了最高达56%(Nahm,2002;Sharpley等,1994;Wodzinski和Wodzinski等,Ullah,1996,如同所充分阐述的,所有这些通过引证结合至本文)。尽管在动物饲料和工业加工中使用植酸酶是有益的,但在动物饲料日粮中使用微生物或植物产生的植酸酶的一个常见的挑战是这些植酸酶在通常用于制备饲料颗粒的调节(conditioning)、挤出或造粒过程中无法保持全部活性。尽管已经设计了一些酶来改善植酸酶的热稳定性,但是大多数酶在造粒期间会失去活性,增加了它们的相对成本并且降低了酶的功效。因此需要具有进一步改善的热稳定性的酶,特别是当饲料制造商更喜欢使用较高温度的造粒工艺时。
本领域众所周知的是,许多生物分子可以通过暴露于高温而变得无活性。由于蛋白质在自然界普遍存在,发生在所有生命界中,并且存在于嗜温菌和嗜热菌多种生物体中,因此它们具有巨大的热稳定性范围。以具有低热稳定性为特征的蛋白质通常通过分子途径进行发展,在分子途径中,它们的结构能量增加,增加了分子振动和运动,这克服了分子内结合力并导致蛋白质展开。当发生展开时,蛋白质内部的结构是无序的,同时暴露蛋白质结构中的亲水和疏水区域以及氨基酸,并且通常导致蛋白聚集。对于具有低热稳定性的蛋白,展开过程通常比重折叠过程快得多,并且在某些情况下可能基本上是不可逆的。相反,具有高度热稳定性的蛋白质通常具有更高程度的分子内键合,在存在增加的热能水平时,这有助于将它们的结构保持在一起,并有助于快速的再折叠率,这可以增强蛋白质在面临不稳定的热暴露时恢复其活性的能力。鉴于在不同的蛋白质之间观察到的热稳定性范围广泛,有机会将不稳定的蛋白质设计为更加热稳定。这与植酸酶特别相关,植酸酶通常来自嗜温或较少的嗜热生物,并且通常很难在动物饲料造粒过程或工业过程中保持高水平的活性。
在植物、微生物细胞或其他细胞生产系统中生产异源蛋白质的另一常见挑战是异源蛋白质作为人类过敏原(变应原)造成的风险。任何异源表达的酶对通过吸入或摄入而暴露于蛋白质的人表现出过敏原风险。为了降低蛋白质,特别是可能不经意地消耗的植物表达的蛋白质的过敏原风险,期望设计植酸酶,使其在暴露于胃环境、肠环境或暴露于胃蛋白酶时,具有降低的稳定性。如果含有工程化植酸酶的植物材料被无意中摄入,则胃蛋白酶稳定性降低使得蛋白质更安全,因为它容易在人体消化道中消化。
发明内容
在一方面中,本发明涉及工程化植酸酶。工程化植酸酶包括目标植酸酶(targetphytase),第一结合元件和第二结合元件。第一结合元件与目标植酸酶融合,并且第二结合元件与目标植酸酶融合。第一结合元件与第二结合元件相互作用以引起工程化植酸酶的环化并且增强目标植酸酶的热稳定性。第一结合元件选自由以下各项组成的组中:内含肽或其部分,卷曲螺旋二聚化结构域或其部分,标签结构域(tag domain)和捕捉结构域(catcher domain)。第二结合元件选自由以下各项组成的组中:标签结构域、捕捉结构域、内含肽或其部分以及卷曲螺旋二聚化结构域或其部分。
在一方面中,本发明涉及编码本文所述的任意一种工程化植酸酶的工程化核酸。
在一方面中,本发明涉及编码工程化植酸酶的工程化核酸。工程化植酸酶包括目标植酸酶,第一结合元件和第二结合元件。第一结合元件和第二结合中的每一个与目标植酸酶融合。第一结合元件与第二结合元件相互作用以引起工程化植酸酶的环化,并增强目标植酸酶的热稳定性。第一结合元件或第二结合元件选自由以下各项组成的组中:标签结构域、捕捉结构域、内含肽或其部分和卷曲螺旋二聚化结构域或其部分。
在一方面中,本发明涉及包括本文所述的任意一种工程化核酸的载体。
在一方面中,本发明涉及包括本文所述的任意一种工程化植酸酶的宿主。宿主选自由以下各项组成的组中:微生物、植物细胞、噬菌体、病毒、哺乳动物细胞和昆虫细胞。
在一方面中,本发明涉及增强目标植酸酶的热稳定性的方法。该方法包括生产本文所述的工程化植酸酶中的任意一种。
在一方面中,本发明涉及制备动物饲料的方法,包括将本文所述的任意一种工程化植酸酶添加至动物饲料。
在一方面中,本发明涉及包括本文所述的任意一种工程化植酸酶的动物饲料。
附图说明
当结合附图阅读时,将更好地理解本发明的优选实施方式的以下详细描述。为了说明本发明的目的,在附图中示出了具体的实施方式。然而,应该理解的是,本发明不限于所示的精确设置和手段。在附图中:
图1是示出了以下的示意图:具有连接至植酸酶编码序列的末端的断裂内含肽的工程化植酸酶(A),断裂内含肽的结合使用非共价结合以环化植酸酶(B),以及在内含肽剪接和共价键形成后产生的环化植酸酶形式(C)。
图2是示出了以下的示意图:具有连接至连接子(该连接子连接至植酸酶编码序列的末端)的断裂内含肽的工程化植酸酶(A),断裂内含肽的结合使用非共价结合以环化植酸酶(B),以及在内含肽的剪接和共价键的形成之后产生的环化植酸酶的形式(C)。
图3是示出了以下的示意图:具有卷曲螺旋结构域(所述卷曲螺旋结构域连接至植酸酶编码序列的末端)的工程化植酸酶(A),卷曲螺旋结构域的结合使用非共价结合以环化植酸酶(B)。
图4是示出了以下的示意图:具有分别与植酸酶编码序列的氨基端和羧基端连接的标签结构域和捕捉结构域的工程化植酸酶(A),标签和捕捉结构域的结合使用非共价结合以环化植酸酶(B),以及在标签-捕捉结构域反应以形成共价键后产生的环化植酸酶的形式(C)。
图5是示出了以下的示意图:具有分别与植酸酶编码序列的羧基端和氨基端连接的标签结构域和捕捉结构域的工程化植酸酶(A),标签和捕捉结构域的结合使用非共价结合以环化植酸酶(B),以及在标签-捕捉结构域反应以形成共价键后产生的环化植酸酶的形式(C)。
图6是示出了以下的示意图:具有连接至连接子(分别连接至植酸酶编码序列的氨基端和羧基端)的标签结构域和捕捉结构域的工程化植酸酶(A),标签结构域和捕捉结构域的结合使用非共价结合以环化植酸酶(B),以及在标签-捕捉结构域反应以形成共价键后产生的环化植酸酶的形式(C)。
图7是示出了以下的示意图:具有连接至连接子(分别连接至植酸酶编码序列的羧基端和氨基端)的标签结构域和捕捉结构域的工程化植酸酶(A),标签结构域和捕捉结构域的结合使用非共价结合以环化植酸酶(B),以及在标签-捕捉结构域反应以形成共价键后产生的环化植酸酶的形式(C)。
图8是示出了表达载体pAG4918的示意图。
图9A-9C是示出了具有与植酸酶编码序列的末端连接的断裂内含肽的所选择的工程化植酸酶的表达盒的示意图。图9A示出了ZmZ27P:xGZein27ss:Gp41-1C:Phy02opt:Gp41-1N:DPNGSEKDEL:NosT。图9B示出了ZmZ27P:Ssp DnaE-C:Phy02opt:Ssp DnaE-N:NosT。图9C示出了ZmZ27P:xGZein27ss:Ssp DnaE-C:Phy02opt:Ssp DnaE-N:DPNGSEKDEL:NosT。
图10A-10H是示出了具有连接至连接子(其连接至植酸酶编码序列末端)的断裂内含肽的所选择的工程化植酸酶的表达盒的示意图。图10A示出了ZmZ27P:Ssp DnaE-C:L33-1:Phy02opt:L33-2:Ssp DnaE-N:NosT。图10B示出了ZmZ27P:xGZein27ss:Ssp DnaE-C:L33-1:Phy02opt:L33-2:Ssp DnaE-N:DPNGSEKDEL:NosT。图10C示出了ZmZ27P:Ssp DnaE-C:L38-1:Phy02opt:L38-2:Ssp DnaE-N:NosT。图10D示出了ZmZ27P:xGZein27ss:Ssp DnaE-C:L38-1:Phy02opt:L38-2:Ssp DnaE-N:DPNGSEKDEL:NosT。图10E示出了ZmZ27P:Ssp DnaE-C:L46-1:Phy02opt:L46-2:Ssp DnaE-N:NosT。图10F示出了ZmZ27P:xGZein27ss:Ssp DnaE-C:L46-1:Phy02opt:L46-2:Ssp DnaE-N:DPNGSEKDEL:NosT。图10G示出了ZmZ27P:Ssp DnaE-C:L55-1:Phy02opt:L55-2:Ssp DnaE-N:NosT。图10H举例示出了ZmZ27P:xGZein27ss:Ssp DnaE-C:L55-1:Phy02opt:L55-2:Ssp DnaE-N:DPNGSEKDEL:NosT。
图11是示出了SspDnaE-C:Phy02:SspDnaE-N构建体的表达谱的凝胶图。
图12是示出了Phy02的热稳定性测定的图。
图13A-13B是示出了SspDnaE-C:Phy02:SspDnaE-N构建体的热稳定性的柱状图。图13A示出了未处理(37℃)和热处理(75℃/60秒)的样品的酶活性。图13B示出了以其各自未处理对照(37℃)的活性的百分比表示的热预处理样品中残留的植酸酶活性。
图14是示出了SpyTag:Phy02:Spy捕捉子(catcher)野生型和突变形式的表达谱的凝胶图。
图15A-15B是示出了SpyTag:Phy02:Spy捕捉子提高了植酸酶的耐热性的柱状图。图15A示出了热预处理的样品的植酸酶活性。图15B示出了热预处理的样品的植酸酶活性的保留。
图16是示出了与野生型酶Phy02(垂直标记)和空载体(水平标记)相比,环状植酸酶gp41-1C:连接子55-1:Phy02:连接子55-2:gp41-1N(闭环)和TrxH:DPNG:gp41-1C[MTT]:连接子55-1:Phy02:连接子55-2:gp41-1N(闭环)的热预处理的图。
图17是示出了在85℃预处理1分钟后,剪接有效的(enabled)和剪接无效的(disabled)(内含肽N125A和连接子S1A)环状植酸酶gp41-1C:连接子55-1:Phy02:连接子55-2:gp41-1N和TrxH:DPNG:gp41-1C[MTT]:连接子55-1:Phy02:连接子55-2:gp41-1N和野生型Phy02植酸酶的植酸酶活性的柱状图。
具体实施方式
仅为了方便而在以下描述中使用某些术语,其并不是限制性的。
如本文所用的,“变体”是指保留与原始序列的生物学活性相同或基本相似的生物学活性的分子。该变体可以来自相同或不同的物种,或者可以是基于天然的分子或现有的分子的合成序列。
一种实施方式包括工程化植酸酶,其包括目标植酸酶、第一结合元件和第二结合元件。第一结合元件可以融合至目标植酸酶,并且第二结合元件可以融合至目标植酸酶。第一结合元件可以与第二结合元件相互作用以引起工程化植酸酶的环化,并且改变目标植酸酶的热稳定性。
第一结合元件和第二结合元件中的每个都能够从工程化植酸酶释放出来。第一结合元件和第二结合元件能够自发地从工程化植酸酶释放出来。当暴露于触发条件时,第一结合元件和第二结合元件能够从工程化植酸酶释放出来。触发条件可以是,但不限于触发温度、触发pH、触发配体结合、触发光、触发离子、离子的触发浓度、触发声音、触发化合物或化合物的触发浓度。
在一种实施方式中,目标植酸酶可以是任意植酸酶。如本文所用,“植酸酶”是能够催化植酸水解的酶。目标植酸酶可以是来源于嗜温、嗜热或超嗜热生物的植酸酶。目标植酸酶可以是来源于真核生物或原核生物的植酸酶。目标植酸酶可以是,但不限于来源于大肠杆菌(Escherichiαcoli)、黑曲霉(Aspergillus niger)、隔孢伏革菌(Peniophora lycii)、粗糙脉孢菌(Neurospora crassa)或意大利拟牛链球菌(Schwαnwmyces accidentαlis)的植酸酶。植酸酶可以被改性以改善热稳定性。当加热到70℃到90℃的温度时,热稳定的植酸酶可以具有活性。在暴露于70℃至90℃的温度时,热稳定的植酸酶可以是活性的。目标植酸酶可以是对胃蛋白酶消化稳定的植酸酶,可以在动物消化道中具有增加的稳定性,并且可以由微生物宿主产生。目标植酸酶可以是易被胃蛋白酶降解的植酸酶。易降解的植酸酶可在胃蛋白酶处理的45分钟至40分钟、40分钟至35分钟、35分钟至30分钟、30分钟至25分钟、25分钟至20分钟、20分钟至15分钟、15分钟至10分钟、10分钟至8分钟、8分钟至6分钟、6分钟至4分钟、4分钟至2分钟的时间段内完全降解。降解的时间段可以在2分钟和45分钟之间的任意两个整数值之间的范围内。所述植酸酶被胃蛋白酶的完全降解可以发生在10分钟内。目标植酸酶可以是商业销售的用于动物饲料的任意植酸酶。
在一种实施方式中,目标植酸酶可以是来源于大肠杆菌的Phy02植酸酶。Phy02植酸酶可以是优化的用于在植物中表达的变体。该变体可以是具有SEQ ID NO:53的氨基酸序列的植酸酶并由密码子优化的核酸序列SEQ ID NO:52编码。该变体可以是具有SEQ ID NO:219的氨基酸序列的植酸酶并且由密码子优化的核酸序列SEQ ID NO:218编码。目标植酸酶可以是具有SEQ ID NO:54的氨基酸序列的Nov9X植酸酶。目标植酸酶可以是具有SEQ IDNO:56的氨基酸序列的CQBscks植酸酶。目标植酸酶可以包括,基本上由或由与选自由SEQID NO:53、54和56组成的组中的参考序列具有具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性。
在一种实施方式中,组合物的植酸酶可以是变体。变体可以包括保守氨基酸取代:即用具有相似性质的氨基酸取代。保守取代可以是极性对极性氨基酸(甘氨酸(G,Gly)、丝氨酸(S,Ser)、苏氨酸(T,Thr)、酪氨酸(Y,Tyr)、半胱氨酸(C,Cys)、天冬酰胺(N,Asn)和谷氨酰胺(Q,Gln));非极性对非极性氨基酸(丙氨酸(A,Ala)、缬氨酸(V,Val)、色氨酸(W,Trp)、亮氨酸(L,Leu)、脯氨酸(P,Pro)、甲硫氨酸(M,Met)、苯丙氨酸(F,Phe));酸性对酸性氨基酸,天冬氨酸(D,Asp)、谷氨酸(E,Glu));碱性对碱性氨基酸(精氨酸(R,Arg)、组氨酸(H,His)、赖氨酸(K,Lys));带电荷氨基酸对带电荷氨基酸,(天冬氨酸(D,Asp)、谷氨酸(E,Glu)、组氨酸(H,His)、赖氨酸(K,Lys)和精氨酸(R,Arg));和疏水对疏水性氨基酸(丙氨酸(A,Ala)、亮氨酸(L,Leu)、异亮氨酸(I,Ile)、缬氨酸(V,Val)、脯氨酸(P,Pro)、苯丙氨酸(F,Phe)、色氨酸(W,Trp)和甲硫氨酸(M,Met))。通过用相同相同氨基酸的不同密码子取代氨基酸的密码子,可以在核酸序列中进行保守性核苷酸的取代。变体可以包括非保守性取代。与未改变的植酸酶相比,变体可具有40%的植酸酶活性。与未改变的植酸酶相比,变体可以具有至少40%、45%、50%、55%、60%、65%、70%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%,或者此处任意两个值之间的整数的活性。植酸酶活性可以通过本文实施例6中描述的比色酶学测定来确定。
在一种实施方式中,具有与相应氨基酸序列SEQ ID NO:53[Phy02]、SEQ ID NO:54[Nov9X]、SEQ ID NO:56[CQBscks]以及SEQ ID NO:219[Phy02opt]具有小于100%的同一性的一种或多种蛋白质是参考蛋白质或氨基酸的变体。在一种实施方式中,提供了分离的蛋白质、多肽、寡肽或肽,沿着SEQ ID NO:53[Phy02]、SEQ ID NO:54[Nov9X]、SEQ ID NO:56[CQBscks]和SEQ ID NO:219[Phy02opt]中任意一条序列的蛋白质的10至50、10至100、10至150、10至300、10至400、10至500、10至600、10至700、10至800、10至900或10至所有氨基酸,所述分离的蛋白质、多肽、寡肽或肽与具有SEQ ID NO:53[Phy02]、SEQ ID NO:54[Nov9X]、SEQ ID NO:56[CQBscks]和SEQ ID NO:219[Phy02opt]中的任意一条序列的蛋白质具有至少70%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性。该序列长度列表涵盖了SEQ ID NO:53[Phy02]、SEQ ID NO:54[Nov9X]、SEQID NO:56[CQBscks]和SEQ ID NO:219[Phy02opt]中的每条全长蛋白质和该列表中的每条更小的长度,甚至涵盖了不超过450个氨基酸的蛋白质。例如,10至50、10至100、10至150、10至300、10至400和10至所有氨基酸的长度将适用于具有400个氨基酸的序列。本文列举的一系列氨基酸序列长度包括该范围内的每条氨基酸序列长度,包括端点在内。所列举的氨基酸长度可以在参考序列内的任意单个位置开始,在该处足够的氨基酸在该单个位置之后以容纳所列举的长度。对于1000个氨基酸或更长的序列,序列长度范围可以通过10-100N氨基酸的增量来扩展,其中N=10或更大的整数。植酸酶的片段可以是本文多肽的子序列,其保留植酸酶的至少40%的活性。该片段可以具有400、405或410个氨基酸。该片段可以包括20、30、40、50、100、150、200、300、400或410个连续的氨基酸。实施方式还包括编码所述氨基酸序列的核酸和识别所述氨基酸序列上的表位的抗体。小于全长氨基酸的序列可以选自以下氨基酸序列中的一条的任意部分:对应于所列举的氨基酸长度的SEQ ID NO:53[Phy02]、SEQ ID NO:54[Nov9X]、SEQ ID NO:56[CQBscks]和SEQ ID NO:NO:219[Phy02opt]。小于全长氨基酸的序列可以选自以下氨基酸序列中的一条的任意部分:SEQ ID NO:53[Phy02]、SEQ ID NO:54[Nov9X]、SEQ ID NO:55[CQBscks]和SEQ ID NO:219[Phy02opt]。
在一种实施方式中,第一结合元件和第二结合元件可以选自由以下各项组成的组中:内含肽或其部分、卷曲螺旋二聚化结构域或其部分,以及标签和捕捉结构域。
在一种实施方式中,第一结合元件或第二结合元件可以是内含肽或其部分。内含肽可以断裂为内含肽部分。断裂内含肽的部分可以来源于嗜热的顺式剪接内含肽。断裂内含肽的部分可以来源于反式剪接内含肽。断裂内含肽的部分可以用于结合植酸酶的末端,并且从而改善其热稳定性。如本文所使用的,术语“断裂内含肽”是指来源于嗜热生物的顺式剪接内含肽,其可以断裂成反式剪接内含肽对或反式剪接内含肽的部分。根据断裂内含肽在分子之间的序列分歧度,断裂内含肽可以通过筛选选自INbase的顺式剪接内含肽来鉴定。对于INbase,参见Perler,F.B(2002).InBase:内含肽数据库,Nucleic acidsresearch,30(1),383-384,如同所充分阐述的,其通过引证结合至本文。这些人工断裂的反式剪接内含肽对可能在N-和C-末端具有规则(canonical)的剪接残基,其中每个新的子结构域将具有至少3.5的净电荷。人工断裂的反式剪接内含肽对可以包括N-内含肽和C-内含肽。N-内含肽可以带正电,C-内含肽可以带负电。可以选择N-内含肽和C-内含肽,目的是当内切核酸酶结构域存在于顺式剪接内含肽前体(从中选择这些断裂内含肽)中时,不将内部的内切核酸酶结构域结合至断裂内含肽组分中。可以基于与微型Tth内含肽(mTth)和GP41-1内含肽的序列比对来选择分割点。这些分割点可以被修饰,并且这些内含肽的变体可以用于本发明。N-内含肽和C-内含肽可以被截短、延伸或修饰,以便在结合植酸酶末端时获得最佳性能,并改善植酸酶消化的热稳定性、表达、溶解性、比活性或胃稳定性。可以将甲硫氨酸残基添加到C-内含肽的氨基末端。
在一种实施方式中,第一结合元件可以是内含肽的C-内含肽并且第二结合元件可以是内含肽的N-内含肽。图1说明C-内含肽可以连接到植酸酶序列的N-末端并且N-内含肽可以连接到植酸酶序列的C-末端。C-内含肽可以是,但不限于Cbu_DnaB-C、Mja_GF6P-C、Mja_Hyp1-C、Mja_IF2-C、Mja_Pol1-C、Pab_CDC211-C、Pab_IF2-C、Pab_VMA-C、Pho_IF2-C、Pho-VMA-C、Rma_DnaB-C、Sru_DnaB-C、Tag_Pol1Tsp-TYPol1-C、Ter_RIR14-C、Tko_IF2-C、Tth-HB27DnaE2-C、Gp41-1C、Gp41-1C[MTT]和Ssp DnaE-C。N-内含肽可以是,但不限于Cbu_DnaB-N、Mja_GF6P-N、Mja_Hyp1-N、Mja_IF2-N、Mja_Pol1-N、Pab_CDC211-N、Pab_IF2-N、Pab_VMA-N、Pho_IF2-N、Pho-VMA-N、Rma_DnaB-N、Sru_DnaB-N、Tag_Pol1Tsp-TYPol1-N、Ter_RIR14-N、Tko_IF2-N、Tth-HB27DnaE2-N、Gp41-1N和SspDnaE-N。C-内含肽可以包括以下氨基酸序列,基本由其组成或由其组成,该氨基酸序列与参考序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%、100%的同一性,所述参考序列选自由以下各项组成的组中:SEQ ID NO:2、4、6、8、10、12、14、16、18、20、22、24、26、28、30、32、189、191和195,并且N-内含肽可以包括以下氨基酸序列,基本上由其组成或由其组成,所述氨基酸序列与参考序列至少具有70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%、100%的同一性,所述参考序列选自由以下各项组成的组中:SEQ ID NO:1、3、5、7、9、11、13、15、17、19、21、23、25、27、29、31、187和193。第一结合元件可以是Cbu_DnaB-C(SEQ ID NO:2),并且第二结合元件可以是Cbu_DnaB-N(SEQ ID NO:1)。第一结合元件可以是Mja_GF6P-C(SEQ ID NO:4),并且第二结合元件可以是Mja_GF6P-N(SEQ ID NO:3)。第一结合元件可以是Mja_Hyp1-C(SEQ ID NO:6),并且第二结合元件可以是Mja_Hyp1-N(SEQ ID NO:5)。第一结合元件可以是Mja_IF2-C(SEQ ID NO:8),并且第二结合元件可以是Mja_IF2-N(SEQ ID NO:7)。第一结合元件可以是Mja_Pol1-C(SEQ ID NO:10),并且第二结合元件可以是Mja_Pol1-N(SEQ ID NO:9)。第一结合元件可以是Pab_CDC211-C(SEQ ID NO:12),并且第二结合元件可以是Pab_CDC211-N(SEQ ID NO:11)。第一结合元件可以是Pab_IF2-C(SEQ ID NO:14),并且第二结合元件可以是Pab_IF2-N(SEQ ID NO:13)。第一结合元件可以是Pab_VMA-C(SEQ ID NO:16),并且第二结合元件可以是Pab_VMA-N(SEQ ID NO:15)。第一结合元件可以是Pho_IF2-C(SEQ ID NO:18),第二结合元件可以是Pho_IF2-N(SEQ ID NO:17)。第一结合元件可以是Pho_VMA-C(SEQ ID NO:20),并且第二结合元件可以是Pho_VMA-N(SEQ ID NO:19)。第一结合元件可以是Rma_DnaB-C(SEQ ID NO:22),并且第二结合元件可以是Rma_DnaB-N(SEQ ID NO:21)。第一结合元件可以是Sru_DnaB-C(SEQ ID NO:24),并且第二结合元件可以是Sru_DnaB-N(SEQ ID NO:23)。第一结合元件可以是Tag_Pol1Tsp-TYPol1-C(SEQ ID NO:26),并且第二结合元件可以是Tag_Pol1Tsp-TYPol1-N(SEQ ID NO:25)。第一结合元件可以是Ter_RIR14-C(SEQ ID NO:28),并且第二结合元件可以是Ter_RIR14-N(SEQ ID NO:27)。第一结合元件可以是Tko_IF2-C(SEQ ID NO:30),并且第二结合元件可以是Tko_IF2-N(SEQ ID NO:29)。第一结合元件可以是Tth-HB27DnaE2-C(SEQ ID NO:32),并且第二结合元件可以是Tth-HB27DnaE2-C(SEQ ID NO:31)。第一结合元件可以是Gp41-1C(SEQ ID NO:189),并且第二结合元件可以是Gp41-1N(SEQ ID NO:187)。第一结合元件可以是Gp41-1C[MTT](SEQ ID NO:191),并且第二结合元件可以是Gp41-1N(SEQ ID NO:187)。第一结合元件可以是Ssp DnaE-C(SEQ IDNO:195),并且第二结合元件可以是Ssp DnaE-N(SEQ ID NO:193)。
在一种实施方式中,第一结合元件和第二结合元件可以是卷曲螺旋二聚化结构域。卷曲螺旋二聚化结构域可以非共价结合目标植酸酶的末端。该卷曲螺旋结构域可以形成稳定的二聚体以结合植酸酶的末端。该卷曲螺旋结构域在长度和序列同一性上可以变化,并且可以被优化以提高给定的表达宿主中的工程化植酸酶的热稳定性、比活性、胃稳定性、胃消化性或异源表达水平。任意的卷曲螺旋结构域可以被用作第一结合元件或第二结合元件以结合植酸酶的末端并由此改善其热稳定性。
在一种实施方式中,第一结合元件可以是卷曲螺旋二聚化结构域的N-螺旋,并且第二结合元件可以是卷曲螺旋二聚化结构域的C-螺旋。图3说明了N-螺旋可以连接至植酸酶序列的N端,并且C-螺旋可以连接至植酸酶序列的C端。N-螺旋可以包括以下氨基酸序列,基本由其组成或由其组成,所述氨基酸序列与参考序列SEQ ID NO:37或39具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性,并且所述C-螺旋可以包括以下氨基酸序列,基本由其组成或由其组成,该氨基酸序列与参考序列SEQ ID NO:38或40具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性。第一结合元件可以是cc17N-端螺旋(SEQ ID NO:37),并且第二结合元件可以是cc17C-端螺旋(SEQ ID NO:38)。第一结合元件可以是cc30N-端螺旋(SEQ ID NO:39),并且第二结合元件可以是cc30C-端螺旋(SEQ ID NO:40)。
在一种实施方式中,第一结合元件或第二结合元件可以是标签结构域或捕捉结构域。标签结构域和捕捉结构域可以结合目标植酸酶的末端并且可以在末端之间产生共价键。标签结构域和捕捉结构域可以有助于目标植酸酶在暴露于高温后的重新折叠,并改善植酸酶的热稳定性。标签结构域和捕捉结构域可以应用于目标植酸酶的C端或N端(如果蛋白质序列重排以促进末端结合,则新形成末端),并且当它们反应时,通常形成稳定的异肽键。
在一种实施方式中,第一结合元件可以是标签结构域或捕捉结构域。第二结合元件可以是标签结构域或捕捉结构域。选为第一结合元件的结构域可以与选为第二个结合元件的结构域不同。图4示出了标签结构域可以连接到植酸酶序列的N端,并且捕捉结构域可以连接到植酸酶序列的C端。图5示出了捕捉结构域可以连接到植酸酶序列的N端,并且标签结构域可以连接到植酸酶序列的C端。标签结构域可以包括以下氨基酸序列,基本上由其组成或由其组成,该氨基酸序列与参考序列SEQ ID NO:33或34具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性。捕捉结构域可以包括以下氨基酸序列,基本上由其组成或由其组成,所述氨基酸序列与参考序列SEQ ID NO:35或36具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性。第一结合元件可以是Phy_catcher1-C(SEQ ID NO:36),并且第二结合元件可以是Phy_tag1-N(SEQ ID NO:33)。第一结合元件可以是Phy_tag1-C(SEQ ID NO:34),并且第二结合元件可以是Phy_catcher1-N(SEQ ID NO:35)。
使用第一结合元件或第二结合元件以进一步促进植酸酶末端的结合,一种实施方式提供了包括一个或多个连接子的工程化植酸酶。一个或多个连接子可以是第一连接子和第二连接子。工程化植酸酶可以包括第一连接子。工程化植酸酶可以包括第二连接子。工程化植酸酶可以包括第一连接子和第二连接子。第一连接子可以与第一结合元件和目标植酸酶邻接,并在它们之间。第二连接子可以与目标植酸酶和第二结合元件邻接,并在它们之间。第一连接子或第二连接子可以是在目标植酸酶与第一结合元件或第二结合元件之间邻接(contiguously)放置的肽序列。当使用断裂内含肽时,断裂内含肽的氨基-内含肽(N-内含肽)和羧基-内含肽(C-内含肽)部分中的一个或两者可连接至第一连接子或第二连接子以及连接至目标植酸酶的末端。在命名连接子时,采用具有前缀“N-”的N-连接子的惯例,其表示N-连接子将连接到期望的结合元件的C-端和植酸酶的N-端。类似地,使用将后缀“-C”附加到C-连接子名称的末尾的惯例,表示C-连接子连接到植酸酶的C-端和期望的结合元件的N-端。
在一种实施方式中,第一连接子可以是N-连接子并且第二连接子可以是C-连接子。例如,图2示出了C-内含肽可以连接至N-连接子(其连接至植酸酶序列的N-末端),并且N-内含肽可连接至C-连接子(其连接至植酸酶序列的C-末端)。图6和图7示出了标签结构域和捕捉结构域通过在植酸酶的氨基端或羧基端使用连接子连接至植酸酶的实例。图6示出了标签结构域可以连接至N-连接子(其连接至植酸酶序列的N端),并且捕捉结构域可以连接至C连接子(其连接至植酸酶序列的C端)。图7示出了捕捉结构域可以连接至N连接子(其连接至植酸酶序列的N端),并且标签结构域可以连接至C连接子(其连接至植酸酶序列的C端)。第一连接子或第二连接子在定位(positioning)第一结合元件或第二结合元件以增强它们的结合,从而提高获得的工程化植酸酶的整体热稳定性上是有用的。根据目标植酸酶和用于设计目标植酸酶的结合元件,第一连接子或第二连接子的长度(定义为至少一个氨基酸长度)、柔性或刚性、等电点结构、疏水性和序列可以变化。相对于使用相同结合元件但缺乏第一连接子或第二结合子的工程化植酸酶,第一连接子或第二连接子或两者可以用于改善工程化植酸酶的热稳定性、表达水平、胃蛋白酶消化率、胃蛋白酶稳定性或比活性。
还可以使用第一连接子或第二连接子的变体。第一连接子或第二连接子可以最初用在工程化植酸酶中,并且随后可以取代氨基酸以提高工程化植酸酶的热稳定性、表达水平、比活性、胃蛋白酶稳定性或胃蛋白酶消化率。第一连接子或第二连接子可以是高度柔性的,并且大部分是非结构化的肽序列。第一连接子或第二连接子可以是刚性的。第一连接子或第二连接子可以形成有序结构。有序结构可以是,但不限于螺旋或卷曲、β-折叠(beta-sheets)或其他结构域。第一连接子或第二连接子可以包括减缓酶展开的速率或提高酶暴露于较高温度后重折叠速率的结构域。第一连接子或第二连接子可以包括增加工程化植酸酶的热稳定性的结构域或结构。第一连接子或第二连接子可以包括另一种酶或具有酶活性的肽序列。
通过包括定点诱变、缺失、插入或其他方法的诱变技术,第一连接子或第二连接子可以容易地被修饰和优化,以用于任意特定目标植酸酶和分子结构的性能。第一连接子或第二连接子的变体可以通过将序列中的氨基酸从N-连接子的N-末端移到C-连接子的C-末端,或者从C-连接子的C-末端的移动到N-连接子的N-末端进行构建。第一连接子或第二连接子可以用于将内含肽分子结构连接至植酸酶。如果需要内含肽剪接,在大多数情况下N-连接子的N-末端必须是丝氨酸、苏氨酸或半胱氨酸氨基酸残基,以便于促进内含肽剪接。此外,已知的是,一些内含肽具有优选的插入位点基序,并且当将这些连接子与给定的内含肽一起使用时,将天然插入位点基序或优选的插入位点基序整合到连接子中可能是有益的。参见Apgar等,2012,A predictive model of intein insertion site for use in theengineering of molecular switches,PloS one,7(5),e37355,如同所充分阐述的,其通过引证结合至本文。
在一种实施方式中,所述第一连接子可以包括以下序列,基本上由其组成或由其组成,所述序列与选自由SEQ ID NO:41、43、45、47、48、50和51组成的组中的序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99或100%的同一性,并且第二连接子可以包括以下序列,基本上由其组成或由其组成,所述序列与选自由SEQ ID NO:42、44、46、49、50和51组成的组中的序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性。第一连接子可以是L33-1连接子(N-连接子)(SEQ ID NO:41),并且第二连接子可以是L33-2连接子(C-连接子)(SEQ ID NO:42)。第一连接子可以是L38-1连接子(N-连接子)(SEQ ID NO:43),并且第二连接子可以是L38-2连接子(C-连接子)(SEQ ID NO:44)。第一连接子可以是L46-1连接子(N-连接子)(SEQ ID NO:45),并且第二连接子可以是L46-2连接子(C-连接子)(SEQ ID NO:46)。第一连接子可以是L55-1.1连接子(N-连接子)(SEQ ID NO:47),并且第二连接子可以是L55-2连接子(C-连接子)(SEQ ID NO:49)。第一连接子可以是L55-1连接子(N-连接子)(SEQ ID NO:48),并且第二连接子可以是L55-2连接子(C-连接子)(SEQ ID NO:49)。第一连接子可以是Phy_taglink(N-连接子)(SEQ ID NO:50),并且第二连接子可以是Phy_catcherlink(C-连接子)(SEQ ID NO:51)。第一连接子可以是Phy_catcherlink(N-连接子)(SEQ ID NO:51),并且第二连接子可以是Phy_taglink(C-连接子)(SEQ ID NO:50)。工程植酸酶的热稳定性可以增强。植酸酶活性可以在70℃至90℃的温度范围内稳定。温度可以是70℃、75℃、80℃、85℃、90℃、70℃至75℃、70℃至80℃、70℃至85℃、70℃至90℃或低于90℃。为了热稳定性而修饰的工程化植酸酶可以通过标准分子生物学技术制备,然后进行筛选。工程化植酸酶可能会发生突变,然后筛选热稳定性。可以使用的筛选系统可以包括λ噬菌体、酵母或允许蛋白质生产和/或其物理和/或功能特性测试的其他表达系统。从工程化蛋白质群体中,可以分离候选物并且可以进一步分析。进一步的分析可以包括DNA测序、功能测定、结构测定、酶活性测定、以及监测热稳定性的变化或响应升高的温度条件的结构。
在一种实施方式中,工程化植酸酶可以包括以下氨基酸序列,基本上由其组成或由其组成,所述氨基酸序列与选自由以下各项组成的组中的参考序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性:SEQ ID NO:58[Cbu_DnaB-C:Phy02:Cbu_DnaB-N(#12Phy02C)]、SEQ ID NO:60[Mja_GF6P-C:Phy02:Mja_GF6P-N(#44Phy02C)]、SEQ ID NO:62[Mja_Hyp1S-N:Phy02:Mja_Hyp1S-C(#46Phy02C)]、SEQ ID NO:64[Mja_IF2-N:Phy02:Mja_IF2-SEQ ID NO:66[Mja_Pol1-C:Phy02:Mja_Pol1-N(#50Phy02C)]、SEQ ID NO:68[Pab_CDC211-C:Phy02:Pab_CDC211-N(#79Phy02C)SEQ ID NO:70[Pab_IF2-C:Phy02:Pab_IF2-N(#81Phy02C)]、SEQ ID NO:72[Pab_VMA-C:Phy02:Pab_VMA-N(#92Phy02C)],SEQ ID NO:74[Pho_IF2–C:Phy02:Pho_IF2-N(#103Phy02C)]、SEQ ID NO:76[Pho_VMA-C:Phy02:Pho_VMA-N(#110Phy02C)]、SEQ ID NO:78[Rma_DnaB-Phy02:Rma_DnaB-N(#116Phy02C)]、SEQ ID NO:80[Sru_DnaB-C:Phy02:Sru_DnaB-N(#123Phy02C)]SEQ ID NO:82[Tag_Pol1_TspTYPol1-C:Phy02:Tag_Pol1_TspTYPol1-N(#128Phy02C)]、SEQ ID NO:84[Ter_RIR14-C:Phy02:Ter_RIR4-N(#135Phy02C)]、SEQ ID NO:86[Tko_IF2-C:Phy02:Tko_IF-N(#143Phy02C)]、SEQ ID NO:88[Tth-HB27_DnaE2–C:Phy02:Tth-HB27_DnaE2–N(#150Phy02C)]、SEQ ID NO:90[Ssp_DnaE–C:Phy02:Ssp_DnaE-N(#225Phy02C)]、SEQ ID NO:92[Gp411–C:Phy02:Gp411-N(#230Phy02C)]、SEQ ID NO:93[Gp411-C:P hy02r14:Gp411-N]、SEQ ID NO:95[Phy02C-27:SspDnaE(SSp_DnaE-C:L33-1:Phy02:L33-2:Ssp_DnaE-N)]、SEQ ID NO:97[Phy02C-32:SspDnaE(SSp_DnaE-C:L38-1:Phy02:L38-2:Ssp_DnaE-N)]、SEQ ID NO:99[Phy02C-40:SspDnaE(SSp_DnaE-C:L46-1:Phy02:L46-2:Ssp_DnaE-N)]、SEQ ID NO:101[Phy02C-49:SspDnaE(SSp_DnaE-C:L55-1:Phy02:L55-2:Ssp DnaE-N)]、SEQ ID NO:103[Phy02-33:cc17(cc17-N:L33-1-Phy02-L33-2:cc17-C)]、SEQ ID NO:105[Phy02-38:cc17(cc17-N:L38-1-Phy02-L38-2:cc17-C)]、SEQ ID NO:107[Phy02-46:cc17(cc17-N:L46-1-Phy02-L46-2:cc17-C)]、SEQ ID NO:109[Phy02-55:cc17(cc17-N:L55-1-Phy02-L55-2:cc17-C)]、SEQ IDNO:111[Phy02-33:cc30(cc30-N:L33-1-Phy02-L33-2:cc30-C)]、SEQ ID NO:113[Phy02-38:cc30(cc30-N:L38-1-Phy02-L38-2:cc30-C)]、SEQ ID NO:115[Phy02-46:cc30(cc30-N:L46-1-Phy02-L46-2:cc30-C)]、SEQ ID NO:117[Phy02-55:cc30(cc30-N:L55-1-Phy02-L55-2:cc30-C)]、SEQ ID NO:119[Tag-Domain:Taglink1:Phy02:Catcherlink1:Catcher]、SEQ ID NO:201[gp41-1C:L55-1:Phy02:L55-2:gp41-1N(#1gp41-Phy02)]、SEQ ID NO:203[gp41-1C[MTT]:L55-1:Phy02:L55-2:gp41-1N(#2gp41-Phy02)]、SEQ ID NO:205[TrxH:DPNG:gp41-1C[MTT]:L55-1:Phy02:L55-2:gp41-1N(#1TrxH-Phy02)]、和SEQ ID NO:207[TrxH:DPNG:gp41-1C[MTT]:L46-1:Phy02:L46-2:gp41-1N(#2TrxH-Phy02)]。
确定两个氨基酸序列或两个核酸序列的百分比同一性可以包括比对和比较两个序列中相应位置处的氨基酸残基或核苷酸。如果两个序列中的所有位置被相同的氨基酸残基或核苷酸占据,那么所述序列被认为是100%相同的。百分比同一性可以通过SmithWaterman算法(Smith TF,Waterman MS 1981“Identification of Common MolecularSubsequences,”Journal of Molecular Biology 147:195-197,如同所充分阐述的,其全部内容通过引证结合)来测量。
在一种实施方式中,提供了编码本文所述的任意一种工程化植酸酶的工程化核酸。编码目标植酸酶的序列可以与选自由以下序列组成的组中的参考序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性:SEQ ID NO:52[Phy02],SEQ ID NO:55[CQBscks],SEQ ID NO:185[Nov9X]和SEQID NO:218[Phy02opt]。
在一种实施方式中,工程化核酸可以包括编码第一结合元件或第二结合元件的序列。工程化核酸可以包括编码内含肽的C-内含肽的序列。工程化核酸可以包括以下序列,基本上由其组成或由其组成,所述序列与以下各序列具有至少70、72、75、80、85、90、91、92、93、94、95、96、97、98、99或100%的同一性:SEQ ID NO:143[Cbu_DnaB-C]、SEQ ID NO:145[Mja_GF6P-C]、SEQ ID NO:147[Mja_Hyp1-C]、SEQ ID NO:149[Mja_IF2-C]、SEQ ID NO:SEQID NO:151[Pab_CDC211-C]、SEQ ID NO:155[Pab_IF2-C]、SEQ ID NO:157[Pab_VMA-C]、SEQID NO:159[Pho_IF2-SEQ ID NO:161[Pho-VMA-C]、SEQ ID NO:163[Rma_DnaB-C]、SEQ IDNO:165[Sru_DnaB-C]、SEQ ID NO:167[Tag_Pol1Tsp-TYPol1-、SEQ ID NO:169[Ter_RIR14-C]、SEQ ID NO:171[Tko_IF2-C]、SEQ ID NO:173[Tth-HB27DnaE2-C]、SEQ ID NO:188[Gp41-1C]190[Gp41-1C[MTT]]和SEQ ID NO:194[Ssp DnaE-C]。工程化核酸可以包括编码内含肽的N-内含肽的序列。工程化核酸可以包括以下序列,基本上由其组成或由其组成,所述序列与选自由以下序列组成的组中的序列具有至少70、72、75、80、85、90、91、92、93、94、95、96、97、98、99或100%的同一性:SEQ ID NO:142[Cbu_DnaB-N]、SEQ ID NO:144[Mja_GF6P-N]、SEQ ID NO:146[Mja_Hyp1-N]、SEQ ID NO:148[Mja_IF2-N]、SEQ ID NO:150[Mja_Pol1-N]、SEQ ID NO:152[Pab_CDC211-N]、SEQ ID NO:154[Pab_IF2-N]、SEQ ID NO:156[Pab_VMA-N]、SEQ ID NO:158[Pho_IF2-N]、SEQ ID NO:160[Pho-VMA-N]、SEQ ID NO:162[Rma_DnaB-N]、SEQ ID NO:164[Sru_DnaB-N]、SEQ ID NO:166[Tag_Pol1Tsp-TYPol1-N]、SEQ ID NO:168[Ter_RIR14-N]、SEQ ID NO:170[Tko_IF2-N]、SEQ ID NO:172[Tth-HB27DnaE2-N]、SEQ ID NO:186[Gp41-1N]和SEQ ID NO:192[Ssp DnaE-N]。
工程化核酸可以包括编码卷曲螺旋二聚化结构域的N-螺旋的序列。工程化核酸可以包括以下序列,基本上由其组成或由其组成,所述序列与SEQ ID NO:178[cc17N-末端螺旋]或SEQ ID NO:180[cc30N-末端螺旋]的序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性。工程化核酸可以包括编码卷曲螺旋二聚化结构域的C卷曲的序列。工程化核酸可以包括以下序列,基本上由其组成或由其组成,所述序列与SEQ ID NO:179[cc17N-末端螺旋]或SEQ ID NO:181[cc30N-末端螺旋]具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性。
工程化核酸可以包括编码标签结构域的序列。工程化核酸可以包括以下序列,基本上由其组成或由其组成,所述序列与SEQ ID NO:174[Phy_tag1-N]或SEQ ID NO:176[Phy_tag1-C]的序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性。工程化核酸可以包括编码捕捉结构域的序列。工程化核酸可以包括以下序列,基本上由其组成或由其组成,该序列与SEQ ID NO:176[Phy_catcher1-N]或SEQ ID NO:177[Phy_catcher1-C]的序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性。
在一种实施方式中,工程化核酸可以包括编码N-连接子或C-连接子的序列。工程化核酸可以包括与参考序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性,所述参考序列选自由以下序列组成的组中:SEQ ID NO:120[L33-1连接子;N-连接子]、SEQ ID NO:122[L38-1连接子;N-连接子]、SEQ ID NO:124[L46-1连接子;N-连接子]、SEQ ID NO:126[L55-1连接子;N-连接子]和SEQID NO:188[L55-1.1连接子;N-连接子]。工程化核酸可以包括与参考序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%同一性的序列,所述参考序列选自由以下序列组成的组中:SEQ ID NO:121[L33-2连接子;C-连接子]、SEQ ID NO:123[L38-2连接子;C-连接子]、SEQ ID NO:125[L46-2连接子;C-连接子]和SEQ ID NO:127:[L55-2连接子;C-连接子]。工程化核酸可以包括其他连接子的序列。工程化核酸可以包括标签连接子(taglinker)或捕捉连接子(catcherlinker),或者两者的序列。工程化核酸可以包括与以下参考序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99或100%同一性的序列:SEQ ID NO:183[Phy_taglink1]或SEQ ID NO:184[Phy_catcherlink1]。
在一种实施方式中,工程化核酸可以包括与选自由以下序列组成的组中的参考序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%同一性的序列:SEQ ID NO:57[Cbu_DnaB-C:Phy02:Cbu_DnaB-N(#12Phy02C)]、SEQ ID NO:59[Mja_GF6P-C:Phy02:Mja_GF6P-N(#44Phy02C)]、SEQ ID NO:61[Mja_Hyp1S–N:Phy02:Mja_Hyp1S-C(#46Phy02C)]、SEQ ID NO:63[Mja_IF2–N:Phy02:Mja_IF2-C(#47Phy02C)]、SEQ ID NO:65[Mja_Pol1–C:Phy02:Mja_Pol1–N(#50Phy02C)]、SEQ IDNO:67[Pab_CDC211-C:Phy02:Pab_CDC211–N(#79Phy02C)]、SEQ ID NO:69[Pab_IF2-C:Phy02:Pab_IF2-N(#81Phy02C)]、SEQ ID NO:71Pab_VMA–C:Phy02:Pab_VMA-N(#92Phy02C)]、SEQ ID NO:73[Pho_IF2–C:Phy02:Pho_IF2-N(#103Phy02C)]、SEQ ID NO:75[Pho_VMA–C:Phy02:Pho_VMA-N(#110Phy02C)]、SEQ ID NO:77[Rma_DnaB–C:Phy02:Rma_DnaB-N(#116Phy02C)]、SEQ ID NO:79[Sru_DnaB–C:Phy02:Sru_DnaB-N(#123Phy02C)]、SEQID NO:81[Tag_Pol1_TspTYPol1–C:Phy02:Tag_Pol1_TspTYPol1–N(#128Phy02C)]、SEQ IDNO:83[Ter_RIR14–C:Phy02:Ter_RIR4-N(#135Phy02C)]、SEQ ID NO:85[Tko_IF2-C:Phy02:Tko_IF-N(#143Phy02C)]、SEQ ID NO:87[Tth-HB27_DnaE2–C:Phy02:Tth-HB27_DnaE2–N(#150Phy02C)]、SEQ ID NO:89[Ssp_DnaE–C:Phy02:Ssp_DnaE-N(#225Phy02C)]、SEQ ID NO:91[Gp411–C:Phy02:Gp411-N(#230Phy02C)]、SEQ ID NO:94[Phy02C-27:SspDnaE(SSp_DnaE-C:L33-1:Phy02:L33-2:Ssp_DnaE-N)]、SEQ ID NO:96[Phy02C-32:SspDnaE(SSp_DnaE-C:L38-1:Phy02:L38-2:Ssp_DnaE-N)]、SEQ ID NO:98[Phy02C-40:SspDnaE(SSp_DnaE-C:L46-1:Phy02:L46-2:Ssp_DnaE-N)]、SEQ ID NO:100Phy02C-49:SspDnaE(SSp_DnaE-C:L55-1:Phy02:L55-2:Ssp DnaE-N)]、SEQ ID NO:102[Phy02-33:cc17(cc17-N:L33-1-Phy02-L33-2:cc17-C)]、SEQ ID NO:104[Phy02-38:cc17(cc17-N:L38-1-Phy02-L38-2:cc17-C)]、SEQ ID NO:106Phy02-46:cc17(cc17-N:L46-1-Phy02-L46-2:cc17-C)]、SEQ IDNO:108[Phy02-55:cc17(cc17-N:L55-1-Phy02-L55-2:cc17-C)]、SEQ ID NO:110[Phy02-33:cc30(cc30-N:L33-1-Phy02-L33-2:cc30-C)]、SEQ ID NO:112[Phy02-38:cc30(cc30-N:L38-1-Phy02-L38-2:cc30-C)]、SEQ ID NO:114[Phy02-46:cc30(cc30-N:L46-1-Phy02-L46-2:cc30-C)]、SEQ ID NO:116Phy02-55:cc30(cc30-N:L55-1-Phy02-L55-2:cc30-C)]、SEQ ID NO:118[Tag-Domain:Taglink1:Phy02:Catcherlink1:Catcher]、SEQ ID NO:128[ZmZ27P:Gp411C:Phy02opt:Gp411N:NosT(#1Phy02opt)]、SEQ ID NO:129[Z27P:xGZein27ss:Gp411-C:Phy02opt:Gp411-N:DPNGSEKDEL:NosT(#2Phy02opt)]、SEQ ID NO:130[ZmZ27P:Ssp_DnaE–C:Phy02opt:Ssp_DnaE–N:N osT(#3Phy02op)t]、SEQ ID NO:131[mZ27P:xGZein27ss:Ssp_DnaE–C:Phy02opt:Ssp_DnaE–N:DPNGSEKDEL:NosT(#4Phy02op)t]、SEQ ID NO:132[ZmZ27P:Ssp_DnaE:L33-1:P hy02opt:L33-2:NosT(SSp_DnaE-C:L33-1:Phy02opt:L33-2:Ssp_DnaE-N)#5Phy02opt、SEQ ID NO:133[ZmZ27P:xGZein27ss:Ssp_DnaE:L33-1:Phy02opt:L33-2:DPNGSEKDEL:NosT(#6Phy02opt])、SEQ ID NO:200[gp41-1C:L55-1:Phy02:L55-2:gp41-1N(#1gp41-Phy02)]、SEQ ID NO:202[gp41-1C[MTT]:L55-1:Phy02:L55-2:gp41-1N(#2gp41-Phy02)]、SEQ ID NO:204[TrxH:DPNG:gp41-1C[MTT]:L55-1:Phy02:L55-2:gp41-1N(#1TrxH-Phy02)]和SEQ ID NO:206[TrxH:DPNG:gp41-1C[MTT]:L46-1:Phy02:L46-2:gp41-1N(#2TrxH-Phy02)]。
工程化核酸可以包括在表达盒中。表达盒可以包括至少一种调控元件。调控元件可以可操作地连接至工程化核酸。在这种情况下,可操作地连接是指调控元件在核酸上赋予其功能。调控元件可以选自:启动子、信号肽、C端延伸和终止子。例如,调控元件可以是启动子,并且可操作地连接启动子将控制工程化核酸的表达。
编码来源于表达盒的工程化植酸酶的工程化核酸的表达可以在启动子(在植物中,提供核酸的转录)的控制之下。启动子可以是组成型启动子或组织特异性启动子或诱导型启动子。组成型启动子可以提供核酸在植物的大部分细胞和组织中以及在多个发育阶段(但不一定是所有阶段)中的转录。只有当暴露于特定的化学或环境刺激时,诱导型启动子才能启动核酸序列的转录。组织特异性启动子能够在特定的植物组织中启动转录。植物组织可以是,但不限于茎、叶、毛状体、花药、玉米芯、种子、胚乳或胚。组成型启动子可以是,但不限于花椰菜花叶病毒(CAMV)35S启动子、夜来香黄色叶卷曲病毒启动子(CMP)、肌动蛋白启动子或二磷酸核酮糖羧化酶小亚基启动子。组织特异性启动子可以是玉米球蛋白启动子(ZmG1b1)、水稻谷蛋白启动子(prGTL)、玉米γ醇溶蛋白(zein)启动子(ZmZ27)或玉米油质蛋白启动子(ZmI1)。信号肽可以是,但不限于玉米γ醇溶蛋白27信号肽或水稻谷蛋白B4信号肽。C端延伸可以是,但不限于HvVSD(来源于大麦(Hordeum vulgare)液泡分选决定簇(Cervelli等人,2004))或SEKDEL(内质网膜定位信号(Endoplasmic reticulum retentionsignal);(Arakawa,Chong,&Langridge,1998;Haq,Mason,Clements,&Arntzen,1995;Korban,2002;Munro&Pelham,1987))。终止子可以是,但不限于NOS(来源于根癌农杆菌(Agrobαcterium tumefaciens)胭脂碱合酶基因)终止子或玉米球蛋白1终止子。
启动子可以是玉米醇溶蛋白27启动子。玉米醇溶蛋白27启动子(ZmZ27P)可以由以下序列编码,所述序列与参考序列SEQ ID NO:137具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性。信号肽可以是玉米醇溶蛋白27信号肽。玉米醇溶蛋白27信号肽(xGZein27ss)可以由以下序列编码,所述序列与参考序列SEQ ID NO:138具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性。C端延伸可以是SEKDEL(SEQ IDNO:140)。SEKDEL可以由与SEQ ID NO:139的序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99或100%同一性的序列编码。终止子可能是NOS终止子。NOS终止子(NosT)可以由与参考序列SEQ ID NO:141具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%同一性的序列编码。
在一种实施方式中,提供了包括本文所述的工程化核酸或表达盒中的任意一种的载体。
本文所述的任意一种工程化植酸酶可以在宿主中表达。宿主可以是,但不限于微生物、植物细胞、噬菌体、病毒、哺乳动物细胞或昆虫细胞。在一种实施方式中,工程化植酸酶的任意一种可以在植物或植物组织中产生。工程化植酸酶可以在将本文所述的工程化核酸中的任意一种或多种导入植物基因组中时产生。工程化核酸可以编码工程化植酸酶或其片段。工程化核酸可以是指导植物表达一种或多种工程化植酸酶的表达盒。将工程化核酸引入植物的方法在本领域中是已知的。该方法可以是用包括编码工程化植酸酶中一种或多种的工程化核酸的载体来转化植物。一种或多种工程化植酸酶可以从植物或植物组织中分离。当暴露于70℃至90℃(包括端点)范围的温度时,本文在转基因植物中表达的一种或多种工程化植酸酶可以具有活性。温度可以是70℃、75℃、80℃、85℃、90℃、70℃至75℃、70℃至80℃、70℃至85℃、70℃至90℃或低于90℃。一种或多种工程化植酸酶可以在任意的转基因植物中生产。
在一种实施方式中,提供了包括本文所述的工程化核酸中任意一种的宿主。宿主可以是,但不限于微生物、植物细胞、噬菌体、病毒、哺乳动物细胞或昆虫细胞。
宿主可以是包括编码本文所述的工程化植酸酶中的任意一种或多种的工程化核酸的转基因植物或其部分。如本文所使用的,转基因植物可指完整的转基因植物或其部分。该部分可以是但不限于叶、茎、花、芽、花瓣、卵巢、果实或种子中的一种或多种。该部分可能是来源于转基因植物的愈伤组织。转基因植物可以由转基因植物的部分再生。转基因植物可以是第一转基因植物和第二转基因植物或非转基因植物的有性杂交的产物,其中产物植物保留引入至第一转基因植物的工程化核酸。一种实施方式提供了本文所述的转基因植物中的任意一种的后代。
在一种实施方式中,提供了增强目标植酸酶热稳定性的方法。改善目标植酸酶热稳定性的一种机制可以是将其N-末端和C-末端以限制末端移动的方式结合在一起。限制末端移动可以增加展开目标植酸酶所需的能量,并且促进目标植酸酶的重新折叠。目标植酸酶末端的结合可以通过分子内共价键和非共价键发生。可以理解的是,结合目标植酸酶的N-和C-末端可以特定地发生在目标植酸酶的第一个氨基酸和目标植酸酶的最后一个氨基酸之间,或者它们之间的任意氨基酸之间的反应中,使得氨基酸之间的反应改善了目标植酸酶的热稳定性。同样地,两个以上的氨基酸可能参与末端的结合,特别是当结合完全或部分使用非共价键时。多种分子内键对于结合目标植酸酶的末端是有用的,所述分子内键包括半胱氨酸键、肽键、异肽键、酰胺键、氢键和其他。因此,该方法可以包括通过将第一结合元件和第二结合元件融合至目标植酸酶来产生工程化植酸酶。在工程化植酸酶中,第一结合元件可以与第二结合元件相互作用。第一结合元件可以与第二结合元件相互作用以引起工程化植酸酶的环化。工程化植酸酶的环化可以改变目标植酸酶的热稳定性。第一结合元件或第二结合元件可以是本文所述的内含肽或其部分、卷曲螺旋二聚化结构域或其部分、标签和捕捉结构域中的任意一种。
工程化的步骤可以包括制备包括编码工程化植酸酶的核酸的表达构建体。
制备表达构建体的步骤可以包括分析对结合目标植酸酶末端有用的分子结构,或者催化反应以在目标植酸酶的末端之间产生共价键。多种分子内键(包括半胱氨酸键、肽键、异肽键、酰胺键、氢键等)对于结合蛋白质的末端是有用的。工程化的步骤可以包括选择可以用于促进植酸酶分子内的共价键或非共价键或两者的形成以提高其热稳定性的分子结构。这些结构可以包括内含肽、标签和捕捉结构域、卷曲螺旋结构域和其他亲和结构域。参见Perler等,1994,Protein splicing elements:inteins and exteins--a definitionof terms and recommended nomenclature.Nucleic acids research,22(7),1125;Gogarten等人,2002,Inteins:structure,function,and evolution.Annual Reviews inMicrobiology,56(1),263-287;Perler,2002,InBase:the intein database.Nucleicacids research,30(1),383-384;Schoene等人,2014,SpyTag/SpyCatchercyclizationconfers resilience to boiling on a mesophilic enzyme.Angewandte ChemieInternational Edition,53(24),6101-6104;Zakeri等,2012,Peptide tag to a rapidcovalent bond to a protein,through engineering a bacterial adhesin。Proceedings of the National Academy of Sciences,109(12),E690-E69;美国申请14/774,954“Use of Dimerization Domains for Temperature Regulation of EnzymeActivity”,如同所充分阐述的,所有这些通过引证结合至本文。可以评估分子结构在结合植酸酶末端上的能力,以及在沿植酸酶末端或靠近末端的点形成共价或非共价键上的能力。分子结构可以用作本文所述方法中的第一结合元件或第二结合元件。分子结构可以是连接到目标植酸酶末端的裂解内含肽,其可以将其氨基-内含肽和羧基-内含肽组分结合在一起,有效地结合植酸酶的末端,但可以不发生形成异肽键或肽键的反应。同样地,在一些情况下,内含肽可以反应形成异肽键或肽键,在后一种情况下,释放与植酸酶结合的内含肽片段并留下完全环化的植酸酶。在这些情况中的每一种情况下,可以在工程化之前,测试工程化植酸酶相对于植酸酶形式在热稳定性上的改善。
制备表达构建体的步骤可以包括对编码工程化植酸酶的序列进行变异。工程化植酸酶的变体可以被构建、筛选并进一步发展。本领域已知多种用于修饰DNA序列和它们所编码的相应蛋白质序列的技术。在这方面可能有用的诱变技术包括定点诱变、饱和诱变(其中在蛋白质序列中的每个位置的每个氨基酸被单独取代,并且选择和组合改进的变体)、随机诱变、结构域互换或交换(domain swapping or exchange)等。另外,当对序列的热稳定性、比活性、宿主表达、胃稳定性或胃消化率进行优化时,小的缺失或插入可能是有益的。
该方法可进一步包括将编码第一结合元件或第二结合元件的核酸与编码目标植酸酶末端的核酸在这样的位置连接,该位置影响结合元件的相互作用并引起目标植酸酶的环化。结合元件可以是断裂内含肽的部分。第一结合元件可以是内含肽的C-内含肽。第二结合元件可以是内含肽的N-内含肽。图1示出了当C-内含肽与植酸酶的N-末端融合,并且N-内含肽与植酸酶的C末端(A结构)融合时,C-内含肽与N-内含肽结合(B结构)。图1还示出了在结合之后,内含肽剪接并且植酸酶的末端通过共价键得到连接(C结构),以及植酸酶被环化。结构C中所示的环化的植酸酶与结构A或B中所示的植酸酶相比可具有增强的热稳定性。图1中示出的结构B是具有C-和N-内含肽结合的中间结构。没有剪接的结合可以稳定工程化植酸酶。然而,这种稳定作用可能不是永久性的,并可能在解离温度下丧失。另一方面,当C-内含肽和N-内含肽的结合进行到剪接时,稳定的共价键可以连接工程化植酸酶的末端,并且可以产生具有高热稳定性的永久性结构(C)。
图2示出了C-内含肽和N-内含肽可以通过连接子连接到植酸酶末端。如图2中所示,N-连接子置于植酸酶的C-内含肽和N-末端之间,并且C-连接子置于植酸酶的N-内含肽和C-末端之间(A结构)。当C-内含肽与N-内含肽结合(B结构)时,内含肽剪接,并且N-连接子通过引起植酸酶环化的共价键连接至C-连接子(C结构)。结构C中所示的环化植酸酶与图2的结构A或B中示出的植酸酶相比,可以具有增强的热稳定性。
结合元件可以是卷曲螺旋二聚化结构域。第一结合元件可以是N-螺旋。第二结合元件可以是C-螺旋。参考图3,N-螺旋和C-螺旋二聚化结构域可以与目标蛋白的N端和C端融合(A结构)。当结构域结合时,植酸酶与结合的结构域一起形成的结构(B结构)与图3中示出的A结构相比,具有增强的热稳定性。卷曲螺旋二聚化结构域可以调节(tailored)为在特定温度下解离或在高温下保持稳定结合。卷曲螺旋的稳定性与七肽重复序列(heptadrepeats)的数目以及疏水性和离子残基的正确配对成正比(Lau等人,1984;Woolfson DN,2005;Parry等人,2008,如同所充分阐述的,所有这些文献均通过引证结合至本文中)。较大的螺旋相互作用(interface)可以增加卷曲螺旋的二聚化强度,并且可以用于将目标蛋白稳定在其熔点以上而无需共价连接。
结合元件可以是标签结构域和捕捉结构域。第一结合元件可以是标签结构域。第二结合元件可以是捕捉结构域。图4示出了标签-结构域可以融合至目标植酸酶的N-末端,并且捕捉-结构域可以融合至目标植酸酶的C-末端(A结构)。当结构域结合(B结构)时,它们通过共价键连接并与目标植酸酶一起形成环状结构(C结构),该环状结构与图4的结构中所示的植酸酶相比,具有增强的热稳定性。图5示出了标签结构域和捕捉结构域域是可互换的,并且该捕捉结构域域可以与目标植酸酶的N-末端融合,标签结构域可以与目标植酸酶的C-末端融合。图6和图7示出了标签结构域和捕捉结构域可以通过连接子连接到植酸酶末端。图4-7中示出的环状结构(C结构)与这些图中示出的非环化的目标植酸酶相比,可以具有增强的热稳定性。
工程化步骤还可以包括使宿主与表达构建体接触。表达构建体可以包括本文所述的任意一种工程化核酸。表达构建体可以插入转化载体中。转化载体可以用于转化宿主。转化可以是,但不限于农杆菌介导的转化、用质粒DNA的电穿孔、DNA摄取、基因枪转化、病毒介导的转化或原生质体转化。转化可以是适用于特定宿主的任意其他转化方法。该方法可以包括选择包括工程化核酸并表达嵌合蛋白的宿主细胞。该方法可以包括将宿主细胞再生为多细胞生物体。该方法可以包括使宿主细胞繁殖以获得多个包括工程化核酸并表达工程化植酸酶的宿主细胞。目标植酸酶的热稳定性可以增强。
在一种实施方式中,提供了包括本文所述的任意一种工程化植酸酶的动物饲料。术语“动物饲料”是指任意的食物、饲料、饲料组合物、制剂、添加剂、补充剂或混合物,它们适于并且旨在用于动物的营养和生长。包括在动物饲料中的工程化植酸酶可以在动物的胃肠或瘤胃环境中有活性。动物饲料中包括的工程化植酸酶可以是对胃蛋白酶消化稳定的植酸酶。动物可以是单胃动物。动物可以是反刍动物。单胃动物可以是,但不限于鸡、火鸡、鸭、猪、鱼、猫或狗。反刍动物可以是,但不限于公牛、母牛、绵羊、马或山羊。工程化植酸酶在制备动物饲料后可以是活性的。在青贮期间,饲料暴露的温度可能在20℃至70℃的范围内。在造粒期间暴露饲料的温度可能在70℃至130℃的范围内。工程化植酸酶可以具有改善的热稳定性并且在饲料造粒期间,在暴露于高温后可以保持活性。
在一种实施方式中,动物饲料还可以包括饲料补充剂(feed supplement)。饲料补充剂可以是任意植物材料。植物材料可以是非转基因植物或工程化植物。植物材料可以包括工程化植物或突变植物。植物材料可以是含有淀粉的谷物。植物材料可以是含有纤维的谷物。植物材料可以是化学处理的草料。饲料补充剂可以是矿物质。矿物质可以是微量矿物质。矿物可质以是宏量矿物(macro mineral)。矿物质可以是磷酸岩或磷酸盐。矿物质可以是磷酸钙。饲料补充剂可以是至少一种维生素。至少一种维生素可以是脂溶性维生素。饲料补充剂可以是氨基酸。饲料补充剂可以含有一种或多种外源性酶。一种或多种外源性酶可以包括水解酶。水解酶可以是EC3.4分类下的水解酶的酶。水解酶可以是,但不限于木聚糖酶、甘露聚糖酶、糖酶(carbohydrase)、蛋白酶、肽酶、葡聚糖酶、纤维素酶、脂肪酶、磷脂酶、果胶酶、半乳糖苷酶、漆酶、淀粉酶、半纤维素酶或纤维二糖水解酶。水解酶可以在包括在饲料补充剂中的工程化植物或其部分中表达。饲料补充剂可以包括纯化的水解酶。饲料补充剂可以是,但不限于生长改善添加剂、着色剂、调味剂、稳定剂、石灰岩、硬脂酸、淀粉、糖类、脂肪酸或树胶。着色剂可以是类胡萝卜素。类胡萝卜素可以是,但不限于鸡油菌素(cantaxanthin)、β-胡萝卜素、虾青素或叶黄素。脂肪酸可以是多不饱和脂肪酸。多不饱和脂肪酸可以包括,但不限于花生四烯酸、二十二碳六烯酸(DHA)、二十碳五烯酸(eicosapentaenoic acid,EPA)或γ-亚油酸。植物材料可以是非转基因植物或其部分。所述植物材料可以包括选自由以下各项组成的组中的至少一种组分:大麦、小麦、黑麦、燕麦、玉米、水稻、黑小麦、甜菜(beet)、甜菜(sugar beet)、菠菜、卷心菜、藜麦、玉米粉、玉米粒、玉米油、酒糟、草料(forage)、小麦粉、小麦粒(wheat pellets)、小麦籽粒(wheat grain)、大麦籽粒、大麦粒、大豆粉、大豆油饼、羽扇豆粉(lupin meal)、菜籽粉、高粱籽粒、高粱粒、油菜籽、向日葵籽和棉籽。
饲料补充剂可以包括选自由以下各项组成的组中的至少一种组分:可溶性固体、脂肪和蛭石、石灰岩(limestone)、纯盐(plain salt)、DL-甲硫氨酸、L-赖氨酸、L-苏氨酸、莫能菌素维生素预混物、磷酸二钙、硒预混物、氯化胆碱、氯化钠和矿物质预混物。饲料补充剂可以包括鱼粉、鱼油、骨粉、羽毛粉和动物脂肪。饲料补充剂可以包括酵母或酵母提取物。
在一种实施方式中,提供了制备动物饲料的方法。该方法可以包括通过本文所述的任意一种方法制备本文所述的任意一种工程化植酸酶。
一种实施方式提供了一种生产动物饲料的方法。该方法可以包括将本文所述的任意一种转基因植物或其部分或其后代与植物材料混合。转基因植物可以是转基因植物的后代。工程化核酸可以包括在基因构建体或表达盒中。该方法可以包括制备本文中的任意转基因植物。转基因植物或其后代可以是植物,其中植酸酶水平可以通过本文的方法增加。该方法可以进一步包括将该混合物造粒。该方法可以进一步包括向混合物中添加饲料补充剂。饲料补充剂可以包括至少一种外源酶。该至少一种外源酶可以选自由以下各项组成的组中的水解酶:木聚糖酶、甘露聚糖酶、蛋白酶、葡聚糖酶和纤维素酶。制备动物饲料可以包括将本文的一种或多种转基因植物与任意其他饲料补充剂组合。
具有工程化核酸(其在植物中编码工程化植酸酶)的表达盒可以在所述方法中的任意时间点(any point)表达。工程化的核酸可以在混合植物的步骤之前表达。工程化核酸可以在植物造粒的步骤期间表达。可以诱导表达。一旦核酸表达,与相同遗传背景但缺乏一个或多个表达盒的非转基因工程化植物中的植酸酶的水平相比,转基因植物可以具有提高的工程化植酸酶水平。
工程化植酸酶可被分离、纯化并作为纯植酸酶添加到动物饲料中。工程化植酸酶可以从完整的宿主生物体中分离出来并作为植酸酶组合物添加到动物饲料中。工程化植酸酶可以与其他饲料补充剂混合添加到动物饲料中。包括工程化植酸酶或纯化的工程化植酸酶的转基因植物可以与其他饲料补充剂组合以形成预混合物。
动物饲料可以生产为粉状饲料。动物饲料可以生产为丸粒(pellet)。研磨的饲料(feed stuffs)可以与预混物混合,该预混物包括任意一种含有工程化植酸酶的转基因植物。工程化植酸酶可以是对胃蛋白酶消化稳定的植酸酶。研磨物可以包括本文所述的植物材料和饲料补充剂。该饲料补充剂可以包括一种或多种本文所述的外源酶。酶可以作为液体或固体制剂加入。对于粉状饲料,固体或液体酶制剂可以在混合步骤之前或期间加入。对于粒状饲料,酶制剂可以在造粒步骤之前或之后加入。植酸酶可以包括在预混物中。所述预混物还可以含有维生素和微量矿物质。宏量矿物质可以单独添加至动物饲料中。
在一种实施方式中,提供了增强目标植酸酶的热稳定性的方法。该方法可以包括产生包括编码植酸酶的工程化核酸的转基因植物。该工程化核酸可以包括本文所述序列中的任意一条。当暴露于70℃至90℃范围(包括端点)的温度时,植酸酶可以是热稳定的。当暴露于70℃至90℃范围(包括端点)的温度时,植酸酶可以是热稳定的。当暴露于70℃、75℃、80℃、85℃、90℃、70℃至75℃、70℃至80℃、70℃至85℃、70℃至90℃或低于90℃范围的温度时,植酸酶可以是热稳定的。热稳定的植酸酶可以是对胃蛋白酶消化稳定的植酸酶。
以下目录包括特定的实施方式。然而,该目录不是限制性的,并且不排除本文中另外描述的实施方式或替代的实施方式。
实施方式
1.一种工程化植酸酶,包括目标植酸酶、第一结合元件和第二结合元件,其中所述第一结合元件和所述第二结合中的每个与所述目标植酸酶融合,所述第一结合元件与所述第二结合元件相互作用以引起所述工程化植酸酶的环化,并且增强所述目标植酸酶的热稳定性,其中所述第一结合元件或所述第二结合元件中的每个选自由以下各项组成的组中:标签结构域、捕捉结构域、内含肽或其部分和卷曲螺旋二聚化结构域或其部分。
2.根据实施方式1所述的工程化植酸酶,其中在所述相互作用时,所述第一结合元件和所述第二结合元件中的每个都能够自发地从所述工程化植酸酶释放出来。
3.根据实施方式1或2中任意一项或两项所述的工程化植酸酶,其中在相互作用时,第一结合元件和第二结合元件中的每个在暴露于触发条件时能够从工程化植酸酶释放出来。
4.根据实施方式3所述的工程化植酸酶,其中所述触发条件选自由触发温度、触发pH、触发配体结合、触发光、触发离子、离子的触发浓度、触发声音、触发化合物或化合物的触发浓度组成的组中。
5.根据前述实施方式中任意一项或多项所述的工程化植酸酶,其中所述第一结合元件或所述第二结合元件与目标植酸酶的N-末端或C-末端融合。
6.根据前述实施方式中任意一项或多项所述的工程化植酸酶,其中所述第二结合元件的N-末端与所述目标植酸酶的C-末端连接并邻接。
7.根据前述实施方式中任意一项或多项所述的工程化植酸酶,其中所述第一结合元件的C末端与所述目标植酸酶的N末端连接并邻接,并且所述第二结合元件的N末端与目标植酸酶的C-末端连接并邻接。
8.根据前述实施方式中任意一项或多项所述的工程化植酸酶,其中所述目标植酸酶选自由以下各项组成的组中:来源于自大肠杆菌(Escherichia coli)、黑曲霉(Aspergillus niger)、隔孢伏革菌(Peniophora lycii)、粗糙脉孢菌(Neurosporacrassa)和意大利拟牛链球菌(Schwaniomyces accidentalis)的植酸酶。
9.根据前述实施方式中任意一项或多项所述的工程化植酸酶,其中所述目标植酸酶包括以下氨基酸序列,该氨基酸序列与选自由SEQ ID NO:53-54、56和219组成的组中的参考序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性。
10.根据前述实施方式中任意一项或多项所述的工程化植酸酶,其中所述第一结合元件是内含肽的C-内含肽,并且所述第二结合元件是内含肽的N-内含肽。
11.根据前述实施方式中任意一项或多项所述的工程化植酸酶,其中所述C-内含肽包括与选自由以下序列组成的组中的参考序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性的氨基酸序列:SEQ ID NO:2、4、6、8、10、12、14、16、18、20、22、24、26、28、30、32、189、191和195;并且所述N-内含肽包括与选自由以下序列组的组中的参考序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性的氨基酸序列:SEQ ID NO:1、3、5、7、9、11、13、15、17、19、21、23、25、27、29、31、187和193。
12.根据实施方式1-9中任意一项或多项所述的工程化植酸酶,其中所述第一结合元件是所述卷曲螺旋二聚化结构域的C-螺旋,并且所述第二结合元件是卷曲螺旋二聚化结构域的N-螺旋。
13.根据实施方式12所述的工程化植酸酶,其中C-螺旋包括与参考序列SEQ IDNO:38或40具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性的氨基酸序列,并且N-螺旋包括与参考序列SEQ ID NO:37或39具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性的氨基酸序列。
14.根据实施方式1-9中任意一项或多项所述的工程化植酸酶,其中第一结合元件和第二结合元件中的每个均包括标签结构域或捕捉结构域,其中选择作为第一结合元件的结构域不同于选择作为第二结合元件的结构域。
15.根据实施方式14所述的工程化植酸酶,其中所述标签结构域包括与参考序列SEQ ID NO:33或34具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性的氨基酸序列。
16.根据实施方式14所述的工程化植酸酶,其中所述捕捉结构域包括与参考序列SEQ ID NO:35或36具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性的氨基酸序列。
17.根据前述实施方式中的任意一个或多个所述的工程化植酸酶,还包括第一连接子和第二连接子,其中所述第一连接子与所述第一结合元件和所述目标植酸酶邻接并且位于它们之间,并且所述第二连接子与所述目标植酸酶和所述第二结合元件邻接并且位于它们之间。
18.根据实施方式17所述的工程化植酸酶,其中第一连接子包括与选自由以下序列组成的组中的参考序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性的序列:SEQ ID NO:41、43、45、47、48、50和51;并且第二连接子包括与选自由以下序列组成的组中的参考序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性的序列:SEQ ID NO:42、44、46、49、50和51。
19.根据前述实施方式中任意一项或多项所述的工程化植酸酶,包括与选自由以下序列组成的组中的参考序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性的氨基酸序列:SEQ ID NO:58、60、62、64、66、68、70、72、74、76、78、80、82、84、86、88、90、92、93、95、97、99、101、103、105、107、109、111、113、115、117、119、201、203、205和207。
20.根据前述实施方式中任意一项或多项所述的工程化植酸酶,其中植酸酶活性在70℃至90℃范围的温度下是稳定的。
21.根据前述实施方式中任意一项或多项所述的工程化植酸酶,其中所述工程化植酸酶在选自由以下各项组成的组中的宿主中表达:微生物、植物细胞、噬菌体、病毒、哺乳动物细胞和昆虫细胞。
22.编码前述实施方式中任意一项或多项所述的工程化植酸酶的工程化核酸。
23.编码工程化植酸酶的工程化核酸,工程化植酸酶包括目标植酸酶、第一结合元件和第二结合元件,其中第一结合元件和第二结合中的每个与目标植酸酶融合,第一结合元件与第二结合元件相互作用以引起所述工程化植酸酶的环化,并且提高所述目标植酸酶的热稳定性,并且所述第一结合元件和所述第二结合元件中的每个选自由以下各项组成的组中:标签结构域、捕捉结构域、内含肽或其部分、以及卷曲螺旋二聚化结构域或其部分。
24.根据实施方式23所述的工程化核酸,其中在相互作用时,第一结合元件和第二结合元件中的每个都能够自发地从工程化植酸酶释放出来。
25.根据实施方式23或24中任意一项或两项所述的工程化核酸,其中在相互作用时,第一结合元件和第二结合元件中的每个在暴露于触发条件时都能够从工程化植酸酶释放出来。
26.根据实施方式25所述的工程化核酸,其中所述触发条件选自由以下各项组成的组:触发温度、触发pH、触发配体结合、触发光、触发离子、离子的触发浓度、触发声音、触发化合物或化合物的触发浓度。
27.根据实施方式23-26中任意一项或多项所述的工程化核酸,其中所述第一结合元件或所述第二结合元件与所述目标植酸酶的N-末端或C-末端融合。
28.根据实施方式23-27中任意一项或多项所述的工程化核酸,其中所述第二结合元件的N-末端与所述目标植酸酶的C-末端连接并邻接。
29.根据实施方式23-28中任意一项或多项所述的工程化核酸,其中所述第一结合元件的C-末端与所述目标植酸酶的N-末端连接并邻接,并且所述第二结合元件的N-末端与所述目标植酸酶的C-末端连接并邻接。
30.根据实施方式23-29中任意一项或多项所述的工程化核酸,包括编码所述植酸酶的序列,所述目标植酸酶选自由由以下各项组成的组中:来源于自大肠杆菌(Escherichia coli)、黑曲霉(Aspergillus niger)、隔孢伏革菌(Peniophora lycii)、粗糙脉孢菌(Neurospora crassa)和意大利拟牛链球菌(Schwaniomyces accidentalis)的植酸酶。
31.根据根据实施方式23-30中任意一项或多项所述的工程化核酸,包括编码所述目标植酸酶的序列,并且与选自由SEQ ID NO:52、55、185和218组成的组中的参考序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性。
32.根据实施方式23-31中任意一项或多项所述的工程化核酸,包括编码第一结合元件的序列,其中第一结合元件是内含肽的C-内含肽。
33.根据实施方式23-32中任意一项或多项所述的工程化核酸,包括编码第二结合元件的序列,其中第二结合元件是内含肽的N-内含肽。
34.根据实施方式32所述的工程化核酸,包括与选自由以下序列组成的组中的参考序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性的序列:SEQ ID NO:143、145、147、149、151、153、155、157、159、161、163、165、167、169、171、173、188、190和194。
35.根据实施方式33所述的工程化核酸,包括与选自由以下序列组成的组中的参考序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性的序列:SEQ ID NO:142、144、146、148、150、152、154、156、158,160、162、164、166、168、170、172、186和192。
36.根据实施方式23-31中任意一项或多项所述的工程化核酸,包括编码第一结合元件的序列,其中第一结合元件是卷曲螺旋二聚化结构域的C-螺旋。
37.根据实施方式23-31和36中任意一项或多项所述的工程化核酸,包括编码第二结合元件的序列,其中第二结合元件是卷曲螺旋二聚化结构域的N-螺旋。
38.根据实施方式36所述的工程化核酸,包括与参考序列SEQ ID NO:179或181具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性的序列。
39.根据实施方式37所述的工程化核酸,包括与参考序列SEQ ID NO:178或180具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性的序列。
40.根据实施方式23-31中任意一项或多项所述的工程化核酸,包括编码第一结合元件的序列,其中第一结合元件是标签结构域或捕捉结构域。
41.根据根据实施方式23-31和40中任意一项或多项所述的工程化核酸,包括编码第二结合元件的序列,其中所述第二结合元件是标签结构域或捕捉结构域,并且其中选择作为第二结合元件的序列不同于选择作为第一结合元件的序列。
42.根据实施方式40和41中任意一项或两项所述的工程化核酸,包括编码标签结构域的序列,并且该序列与参考序列SEQ ID NO:174或175具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性。
43.根据实施方式40-42中任意一项或多项所述的工程化核酸,包括编码捕捉结构域的序列,并且该序列与参考序列SEQ ID NO:176或177具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性。
44.根据实施方式23-43中任意一项或多项所述的工程化核酸,还包括编码第一连接子的序列和编码第二连接子的序列,其中所述第一连接子与所述第一结合元件和所述目标植酸酶邻接并位于它们之间,所述第二连接子与所述第所述目标植酸酶和二结合元件邻接并位于它们之间。
45.根据实施方式44所述的工程化核酸,包括编码第一连接子的序列,并且该序列与选自由以下序列组成的组中的序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性:SEQ ID NO:120、122、124、126、182、183和184;以及编码第二连接子的序列,并且该序列与选自由以下序列组成的组中的序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性:SEQ ID NO:121、123、125、127、183和184。
46.根据根据实施方式23-45中任意一项或多项所述的工程化核酸,包括与选自由以下序列组成的组中的参考序列具有至少70%、72%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的同一性的序列:SEQ ID NO:57、59、61、63、65、67、69、71、73、75、77、79、81、83、85、87、89、91、94、96、98、100、102、104、106、108、110、112、114、116、118、128-133、200、202、204和206。
47.根据实施方式23-46中任意一项或多项所述的工程化核酸,包括编码在70℃至90℃范围的温度下具有稳定植酸酶活性的工程化植酸酶的序列。
48.根据实施方式23-47中任意一项或多项所述的工程化核酸,所述工程化核酸在选自由微生物、植物细胞、噬菌体、病毒、哺乳动物细胞和昆虫细胞组成的组中的宿主中表达。
49.根据实施方式48所述的工程化核酸,其中所述宿主是植物细胞。
50.一种包括编码实施方式1-21中任意一项或多项所述的工程化植酸酶的工程化核酸的载体。
51.一种包括实施方式23-48中任意一项或多项所述的工程化核酸的载体。
52.一种宿主,包括实施方式1-21中任意一项或多项所述的工程化植酸酶或实施方式23-48中任意一项或多项所述的工程化核酸,其中所述宿主选自由以下各项组成的组中:微生物、植物细胞、噬菌体、病毒、哺乳动物细胞和昆虫细胞。
53.一种增强目标植酸酶的热稳定性的方法,包括生产实施方式1-21中任意一项或多项所述的工程化植酸酶。
54.一种动物饲料,包括实施方式1-21中任意一项或多项所述的工程化植酸酶。
55.根据实施方式54所述的动物饲料,还包括饲料补充剂。
56.根据实施方式55所述的动物饲料,其中所述饲料补充剂是植物材料。
57.根据实施方式56所述的动物饲料,其中植物材料是非转基因植物或工程化植物。
58.根据实施方式54-57中任意一项或多项所述的动物饲料,其中所述饲料补充剂包括一种或多种外源酶。
59.根据实施方式58所述的动物饲料,其中所述一种或多种外源酶包括选自由以下各项组成的组中的水解酶:木聚糖酶、内切葡聚糖酶、纤维素酶、蛋白酶、葡聚糖酶、淀粉酶和甘露聚糖酶。
60.根据实施方式54-59中任意一项或多项所述的动物饲料,其中所述植物材料包括选自由以下各项组成的组中的至少一种组分:玉米粉、玉米粒、小麦粉、小麦粒、小麦籽粒、大麦籽粒、大麦粒、大豆粉、大豆油饼、高粱籽粒和高粱粒。
61.根据实施方式55-60中任意一项或多项所述的动物饲料,其中所述饲料补充剂包括选自由以下各项组成的组中的至少一种组分:可溶性固体、脂肪和蛭石、石灰岩、纯盐、DL-甲硫氨酸、L-赖氨酸、L-苏氨酸、维生素预混物、磷酸二钙、硒预混物、氯化胆碱、氯化钠和矿物质预混物。
62.一种制备动物饲料的方法,其包括将实施方式1-21中任意一项或多项所述的工程化植酸酶添加到动物饲料中。
63.根据实施方式62所述的方法,还包括将所述混合物造粒。
64.根据实施方式62或63中任意一项或两项所述的方法,还包括向所述混合物中添加饲料补充剂。
65.根据实施方式64所述的方法,其中所述饲料补充剂包括至少一种外源酶。
66.根据实施方式65所述的方法,其中所述至少一种外源酶是选自由木聚糖酶、甘露聚糖酶、蛋白酶、葡聚糖酶和纤维素酶组成的组中的水解酶。
67.一种促进从动物中的植酸或植酸盐释放无机磷酸盐的方法,包括用动物饲料喂养动物,所述动物饲料含有实施方式1-22中任意一项或多项的工程化植酸酶。
68.根据实施方式31所述的方法,还包括根据实施方式62-66中任意一项或多项所述的方法制备动物饲料。
69.根据实施方式67或68中任意一项或两项所述的方法,其中所述动物是单胃动物或反刍动物。
70.一种环化植酸酶,包括实施方式1、5-9和12-21中任意一项或多项所述的工程化植酸酶,其中第一结合元件结合至第二结合元件。
71.一种环化植酸酶,包括实施方式1-10和13-21中任意一项或多项所述的工程化植酸酶,其中在相互作用时,所述第一结合元件和所述第二结合元件从所述工程化植酸酶释放出来,并且目标植酸酶的N-末端和目标植酸酶的C-末端连接。
本文中进一步的实施方式可以通过利用来自本文的任意一个或多个其它实施方式的一个或多个要素来补充实施方式形成,和/或利用来自本文的一个或多个其他实施方式的一个或多个元素替换来自一个或多个实施方式的一个或多个要素。
实施例
提供以下非限制性实施例以说明特定的实施方式。通篇的实施方式可以用来自以下的一个或多个实施例的一个或多个细节进行补充,和/或来自实施方式的一个或多个要素可以用来自以下的一个或多个实施例的一个或多个细节替代。
实施例1.用于改善植酸酶热稳定性的遗传元件的描述
用于改善植酸酶热稳定性的分子结构或结构域。在对于结合蛋白质末端并且,或催化在蛋白质末端之间产生共价键的反应有用的分子结构为内含肽和标签结构域和捕捉结构域。
内含肽:尽管在本发明中可以使用任意的断裂内含肽来结合植酸酶的末端并由此改善其热稳定性,但是使用了一组来自嗜热的顺式剪接内含肽的内含肽。根据之间的序列分歧(sequence divergence),该组通过筛选选自INbase的一组157个的顺式剪接内含肽进行聚集。对于INbase,参见Perler,F.B.(2002).InBase:the intein database.Nucleicacids research,30(1),383-384,如同所充分阐述的,其通过引证结合至本文。选择来自嗜热生物的顺式剪接内含肽并将其分成反式剪接内含肽对。这些人工的断裂内含肽需要在N-末端和C-末端具有典型的(canonical)剪接残基,其中每个新的子结构域将具有至少3.5的净电荷。这导致了18个断裂内含肽,其中所有N-内含肽都带正电并且C-内含肽带负电。选择N-末端结构域和C-末端结构域的目的是,当内切核酸酶结构域存在于顺式剪接内含肽前体(从中选择这些断裂内含肽)中时,不将内部的内切核酸酶结构域引入至断裂内含肽组分(即N-内含肽或C-内含肽)。然后根据与微型Tth内含肽(mTth)和GP41-1内含肽的序列比对来选择分割点。将甲硫氨酸残基添加到了以下集合中的C-内含肽的氨基末端。表1中示出的反式剪接内含肽的序列如下:
表1反式剪接内含肽的序列
标签结构域和捕捉结构域:标签结构域和捕捉结构域可以在蛋白质的末端之间产生共价键,并且用于在暴露于高温后帮助蛋白质的重新折叠。表2中示出的标签结构域和捕捉结构域的序列如下。
表2标签-捕捉结构域的序列
氨基酸序列编号 核酸序列编号 序列说明
33 174 Phy_tag1-N
34 175 Phy_tag1-C
35 176 Phy_catcher1-N
36 177 Phy_catcher1-C
卷曲螺旋二聚化结构域:一组卷曲螺旋结构域可以如表3中所描述的使用,并在图3中进行了阐述。在表3中示出了卷曲螺旋结构域的序列。
表3卷曲螺旋结构域的序列
序列编号氨基酸 序列编号核酸 序列说明
37 178 cc17N-末端螺旋
38 179 cc17C-末端螺旋
39 180 cc30N-末端螺旋
40 181 cc30C-末端螺旋
卷曲螺管cc17设计用于热稳定性,在高温下形成二聚体,其在高达至少60℃时稳定。相反的,卷曲螺旋cc30在<30℃的温度下形成二聚体,并在50℃左右的温度下开始解离。
连接子:连接子在序列组成和长度上都有所变化。表4中示出了连接子的序列。
表4连接子的序列
氨基酸序列编号 核酸序列编号 序列说明
41 120 L33-1连接子(N-连接子)
42 121 L33-2连接子(C-连接子)
43 122 L38-1连接子(N-连接子)
44 123 L38-2连接子(C-连接子)
45 124 L46-1连接子(N-连接子)
46 125 L46-2连接子(C-连接子)
47 182 L55-1.1连接子(N-连接子)
48 126 L55-1连接子(N-连接子)
49 127 L55-2连接子(C-连接子)
50 183 Phy_标签连接
51 184 Phy_捕捉连接
199 198 DPNG连接子
一种利用选择的分子结构和任意期望的连接子(如果必要)构建的工程化植酸酶(其具有增加的热稳定性)可以对胃蛋白酶消化稳定,如可能用于微生物产品以增加其在动物中的稳定性,或者它可能容易被胃蛋白酶降解(在不到30分钟或不到10分钟)以降低其潜在的致敏性。
目标植酸酶:虽然任意植酸酶都可以用作本发明的目标植酸酶,但用于在植物中表达的一种目标植酸酶是来源于大肠杆菌的Phy02植酸酶变体。以下给出了不具有信号序列、前导序列或第一个甲硫氨酸的酶的大肠杆菌密码子优化序列(Phy02opt)。
目标植酸酶的序列示出了在表5中。
表5目标植酸酶的序列
实施例2.使用直接连接至植酸酶的内含肽构建工程化植酸酶
使用本领域已知的标准重组DNA和分子生物学技术(Ausubel,Current Methodsin Molecular Biology)构建编码工程化植酸酶或环化植酸酶分子的基因。可替代地,完全合成的基因可以从规定的酶序列的设计中直接定制并获得。这类合成DNA序列可以从供应商处获得,密码子优化以用于在任意特定生物体(微生物、植物、哺乳动物等)中表达,并包括任意可以有利于克隆和表达的期望的限制性位点。
在没有信号序列的情况下,将植酸酶(Phy02,SEQ ID NO.52,作为该实施例中的目标植酸酶,但可以被其他植酸酶替代)的DNA序列与编码反式-剪接内含肽部分的DNA序列融合以在分子的氨基末端构建编码C-内含肽的线性分子,分子的羧基端直接与Phy02植酸酶的氨基末端融合,并且N-内含肽的氨基末端直接与Phy02植酸酶的羧基末端融合(C-内含肽:Phy02:N-内含肽),如图1中所描述的。图1示出了具有与植酸酶编码序列末端连接的断裂内含肽的工程化植酸酶(A),使用非共价结合使断裂内含肽结合以环化植酸酶(B),以及在内含肽剪接和共价键形成后产生的环化植酸酶形式(C)。将构建体克隆到pETDuet I表达载体的EcoRI和XhoI位点之间并转化到Shuffle T7大肠杆菌宿主(NEB)中。本领域技术人员将了解内含肽剪接的要求,并且将理解在目标植酸酶的C-内含肽和氨基末端之间的连接处需要适当的氨基酸以促进内含肽剪接。参见Apgar等人,2012,A predictive model ofintein insertion site for use in the engineering of molecular switches.PloSone,7(5),e37355;Xu,M.Q.,&Perler,F.B.,1996,The mechanism of protein splicingand its modulation by mutation.The EMBO journal,15(19),5146,如同所充分阐述的,两者均通过引证结合至本文。在这个实施例中,这个单一的氨基酸是否被认为是连接子或作为目标植酸酶的一部分,并不是分化(differentiation)的关键点。在该实施例中,在Phy02植酸酶的N末端添加单个丝氨酸氨基酸可以被认为是C-内含肽和目标植酸酶之间的连接子,其长度为一个氨基酸。该单个氨基酸丝氨酸连接子可以被苏氨酸或半胱氨酸取代。以下列出了构建体的核苷酸序列。反式剪接C-内含肽和N-内含肽的核苷酸序列为大写的,剪接必要丝氨酸(agc)被添加到植酸酶序列的N-末端,为黑体字,Phy02植酸酶的序列为小写字母加下划线字符。
编码Cbu_DnaB-C:Phy02:Cbu_DnaB-N(#12Phy02C)[氨基酸(AA)_SEQ ID NO:58]的核苷酸序列如下:
>编码Mja_GF6P-C:Phy02:Mja_GF6P-N(#44Phy02C)[AA_SEQ ID NO:60]的核苷酸序列如下:
>编码Mja_Hyp1S-N:Phy02:Mja_Hyp1S-C(#46Phy02C)[AA_SEQ ID NO:62]的核苷酸序列如下:
>编码Mja_IF2-N:Phy02:Mja_IF2-C(#47Phy02C)[AA_SEQ ID NO:64]的核苷酸序列如下:
>编码Mja_Pol1–C:Phy02:Mja_Pol1–N(#50Phy02C)[AA_SEQ ID NO:66]的核苷酸序列如下:
>编码Pab_CDC211-C:Phy02:Pab_CDC211–N(#79Phy02C)[AA_SEQ ID NO:68]的核苷酸序列如下:
>编码Pab_IF2-C:Phy02:Pab_IF2-N(#81Phy02C)[AA_SEQ ID NO:70]的核苷酸序列如下:
>编码Pab_VMA-C:Phy02:Pab_VMA-N(#92Phy02C)[AA_SEQ ID NO:72]的核苷酸序列如下:
>编码Pho_IF2–C:Phy02:Pho_IF2-N(#103Phy02C)[AA_SEQ ID NO:74]的核苷酸序列如下:
>编码Pho_VMA–C:Phy02:Pho_VMA-N(#110Phy02C)[AA_SEQ ID NO:76]的核苷酸序列如下:
>编码Rma_DnaB‐C:Phy02:Rma_DnaB‐N(#116Phy02C)[AA_SEQ ID NO:78]的核苷酸序列如下:
>编码Sru_DnaB‐C:Phy02:Sru_DnaB‐N(#123Phy02C)[AA_SEQ ID NO:80]的核苷酸序列如下:
>编码Tag_Pol1_TspTYPol1-C:Phy02:Tag_Pol1_TspTYPol1-N(#128Phy02C)[AA_SEQ ID NO:82]的核苷酸序列如下:
编码Ter_RIR14-C:Phy02:Ter_RIR4-N(#135Phy02C)[AA_SEQ ID NO:84]的核苷酸序列如下:
>编码Tko_IF2-C:Phy02:Tko_IF-N(#143Phy02C)[AA_SEQID NO:86]的核苷酸序列如下:
编码Tth-HB27_DnaE2-C:Phy02:Tth-HB27_DnaE2-N(#150Phy02C)[AA_SEQ ID NO:88]的核苷酸序列如下:
编码Ssp_DnaE-C:Phy02:Ssp_DnaE-N(#225Phy02C)[AA_SEQ ID NO:90]的核苷酸序列如下:
>编码Gp411-C:Phy02:Gp411-N(#230Phy02C)[AA_SEQ ID NO:92]的核苷酸序列如下:
本领域技术人员将认识到,可以创建、筛选和进一步开发这些序列的多种变体。本领域已知多有种用于修饰DNA序列和它们编码的相应蛋白质序列的技术。在这方面有用的诱变技术包括定点诱变、饱和诱变(其中在蛋白质序列中的每个位置处每个氨基酸被单独取代,并且选择和组合改进的变体)、随机诱变、结构域互换(swapping)或交换(exchange)等。另外,当优化序列的热稳定性、比活性、宿主表达、胃稳定性或胃消化率时,小的缺失或插入可以是有益的。
在此特定的实施例中,当期望将内含肽直接融合至目标植酸酶的末端而不添加另一丝氨酸氨酸时,因为目标植酸酶序列Phy02(SEQ ID NO:53)以AQSEPELKLE...开始(SEQID NO:134),显而易见的是,在本实施例中提供的每条序列中,如果植酸酶序列的前两个氨基酸丙氨酸和谷氨酰胺(AQ)缺失(导致SEPELKLE...(序列编号:135)),并且所得到的植酸酶序列(SEPELKLE...(序列编号:135))氨基末端的第一个丝氨酸被用作丝氨酸以促进内含肽剪接,在C-内含肽(...HN)的羧基末端与植酸酶(AQSEPELKLE...(序列编号:134))的氨基末端之间所添加的丝氨酸氨基酸(...S...)并不是必须的。如果在末端的结合期间,需要重新组装整个目标植酸酶序列(包括缺失的丙氨酸和谷氨酰胺),从植酸酶序列的氨基末端除去的丙氨酸和谷氨酰胺可以添加到植酸酶序列的羧基末端,正好在与N-内含肽的连接处。通过这种方式,植酸酶的整个天然序列将在内含肽剪接反应后重新组装,而目标植酸酶序列没有明显的重排。同样,即使内含肽结合以环化蛋白质(而不剪接),添加的丙氨酸和谷氨酰胺在空间上的位置将与其留在植酸酶的氨基末端时末端的结合之后其原本的位置相似。
从植酸酶去除氨基末端氨基酸残基并将它们依次添加到羧基末端的这种技术可以延伸并应用于目标植酸酶中任意期望的内含肽插入点。这提供了用于促进基于内含肽的结合和/或目标植酸酶的环化的通用算法和技术。例如,如果目标植酸酶的末端在空间上距离太远,而不能使用内含肽、标签-捕捉结构域、卷曲螺旋结构域或其他分子结构来实现末端的有效结合,则可以通过从氨基末端移动氨基酸,并且将它们依次添加到目标植酸酶的羧基末端,来选择新的一组末端,并且将分子结构添加到新选择的末端。
为了说明上述重排技术,Gp411-C:Phy02:Gp411-N(#230Phy02C)的最终蛋白质序列可以重新排列如下:(Phy02(粗体)氨基酸串AQSEPELKLESVVIV(SEQ ID NO:136)从其N-末端移至其C-末端)。Gp411-C:Phy02r14:Gp411-N的氨基酸序列如下:
实施例3使用与植酸酶连接的内含子创建工程化植酸酶
类似于实施例2,工程化或环化的植酸酶可以使用如图2所示的连接子序列构建。图2示出了具有连接至连接子的断裂内含肽的工程化植酸酶,所述连接子连接至植酸酶编码序列的末端(A),使用非共价结合使断裂内含肽结合以环化植酸酶(B),以及在内含肽剪接和共价键形成后产生的环化植酸酶的形式(C)。如实施例2中描述的,这类分子可以使用已知的重组DNA和分子生物学方法制备,或者通过直接排序编码这些工程化植酸酶的DNA序列来制备。样品连接子序列在实施例1中列出了,并且用于构建以下工程化植酸酶,其中内含肽序列为大写,连接子序列为斜体下划线小写字体,并且植酸酶序列为小写且未斜体。
Phy02C-27:SspDnaE(SSp_DnaE-C:L33-1:Phy02:L33-2:Ssp_DnaE-N)的氨基酸序列及编码其的核苷酸序列如下:
Phy02C-32:SspDnaE(SSp_DnaE-C:L38-1:Phy02:L38-2:Ssp_DnaE-N)的氨基酸序列及编码其的核苷酸序列如下:
Phy02C-40:SspDnaE(SSp_DnaE-C:L46-1:Phy02:L46-2:Ssp_DnaE-N)的氨基酸序列和编码其的核苷酸序列如下:
Phy02C-49:SspDnaE(SSp_DnaE-C:L55-1:Phy02:L55-2:Ssp DnaE-N)的氨基酸序列和编码其的核苷酸序列如下:
这些工程化植酸酶可以使用已知技术与其它分子相同地评估热稳定性、来自任何期望的宿主(微生物、植物或其他)的异源表达水平、比活性、胃稳定性或胃消化率(Thomas,K等,2004,A multi-laboratory evaluation of a common in vitro in pepsindigestion assay protocol used in assessmenting the safety of novelproteins.Regulatory Toxicology and Pharmacology,39(2),87-98;FU,TJ(2002),Digestion stability as a criterion for protein allergenicityassessment.Annals of the New York Academy of Sciences,964(1),99-110,如同所完全阐述的,所有这些文献通过引证并入本文)。
实施例4.使用卷曲螺旋结构域构建工程化植酸酶
基于来自实施例3的工程化植酸酶设计以下分子。这些分子含有连接子,但是反式剪接C-内含肽和N-内含肽分别用N-末端螺旋和C-末端螺旋取代。四种原型设计在连接子长度和组成上不同。
下面是四种原型卷曲螺旋稳定的植酸酶的核苷酸和氨基酸序列。N-末端和C-末端的螺旋序列为大写,连接子序列为小写斜体,植酸酶序列为小写。
编码Phy02-33:cc17(cc17-N:L33-1-Phy02-L33-2:cc17-C)[AA_SEQ ID NO:103]的核苷酸序列如下:
编码Phy02-38:cc17(cc17-N:L38-1-Phy02-L38-2:cc17-C)[AA_SEQ ID NO:105]的核苷酸序列如下:
编码Phy02-46:cc17(cc17-N:L46-1-Phhy02-L46-2:cc17-C)[AA_SEQ ID NO:107]的核苷酸序列如下:
编码Phy02-55:cc17(cc17-N:L55-1-Phy02-L55-2:cc17-C)[AA_SEQ ID NO:109]的核苷酸序列如下:
热不稳定的卷曲螺旋修饰的植酸酶(对照;具有四种原型连接子的cc30)。
编码Phy02-33:cc30(cc30-N:L33-1-Phy02-L33-2:cc30-C)[AA_SEQ ID NO:111]的核苷酸序列如下:
编码Phy02-38:cc30(cc30-N:L38-1-Phy02-L38-2:cc30-C)[AA_SEQ ID NO:113]的核苷酸序列如下:
编码Phy02-46:cc30(cc30-N:L46-1-Phy02-L46-2:cc30-C)[AA_SEQ ID NO:115]的核苷酸序列如下:
编码Phy02-55:cc30(cc30-N:L55-1-Phy02-L55-2:cc30-C)[AA_SEQ ID NO:117]的核苷酸序列如下:
实施例5使用标签-捕捉结构域集合构建工程化植酸酶
如图4-7所描述的,使用实施例1中描述的方法,工程化植酸酶可以使用标签和捕捉结构域构建。图4示出了具有标签结构域和捕捉结构域分别连接至植酸酶编码序列的氨基端和羧基端的工程化植酸酶(A),以及标签结构域和捕捉结构域使用非共价连接结合以环化植酸酶(B),以及在标签-捕捉结构域反应以形成共价键后产生的的环化植酸酶的形式(C)。图5示出了具有标签-结构域和捕捉-结构域分别连接至植酸酶编码序列的羧基端和氨基端的工程化植酸酶(A),以及标签-结构域和捕捉-结构域使用非共价连接结合以环化植酸酶(B),以及在标签-捕捉结构域反应以形成共价键后产生的的环化植酸酶的形式(C)。图6示出了具有标签结构域和捕捉结构域连接至连接子(所述连接子分别连接至植酸酶编码序列的氨基端和羧基端)的工程化植酸酶(A),以及标签结构域和捕捉结构域使用非共价连接结合以环化植酸酶(B),以及在标签-捕捉结构域反应以形成共价键后的产生的环化植酸酶的形式(C)。图7示出了具有标签结构域和捕捉结构域连接至连接子(所述连接子分别连接至植酸酶编码序列的羧基端和氨基端)的工程化植酸酶(A),以及标签结构域和捕捉结构域使用非共价连接结合以环化植酸酶(B),以及在标签-捕捉结构域反应以形成共价键后产生的环化植酸酶的形式(C)。
标签结构域和捕捉结构域可以直接连接至植酸酶的末端,或使用连接子连接至末端。与通常具有优选末端(内含肽的每个部分连接至该优选末端)的断裂内含肽不同,标签结构域和捕捉结构域可以在任意末端使用。例如,一种工程化植酸酶可以具有连接至目标植酸酶的氨基末端的标签结构域,而无连接子(图4)或具有连接子(图6),并且具有连接至目标植酸酶羧基末端的捕捉结构域,而无连接子(图4)或具有连接子(图6)。类似地,一种工程化植酸酶可以具有连接至目标植酸酶羧基末端的标签结构域,而无连接子(图5)或具有连接子(图7),并且具有连接至目标植酸酶氨基末端的捕捉结构域,而无连接子(图5)或具有连接子(图7)。标签结构域和捕捉结构域能够在两种构型中结合目标植酸酶的末端并通过形成共价键形成环状植酸酶。以下序列示出了如何构建工程化Phy02植酸酶:
标签结构域:Tlinker1:Phy02:Clinker1:Catcher(连接子用粗体和下划线表示):
如同本文所描述的其他工程化分子,可以使用本文描述的分子和分子变体的优化及方法。可以采用多种不同的优化和诱变方法,如实施例2和3以及本说明书其他地方所描述的。
本领域技术人员还将认识到,具有不同分子结构和结合结构域的任意目标植酸酶都可以用于上述任意实施例中。例如,标签结构域和捕捉结构域可以连接到CQBscks植酸酶上,有或没有连接子,以构建具有改善的热稳定性的植酸酶版本。同样,任意其他结构(包括内含肽和卷曲螺旋)可以与CQBscks或任意其他目标植酸酶一起使用,以改善目标植酸酶的热稳定性。
实施例6植酸酶活性的测定
如本文所述的,植酸酶测定对于工程化植酸酶的改善的热稳定性是必需的。参见Engelen等,2001,Determination of phytase activity in feed by a colorimetricenzymatic method:collaborative interlaboratory study.Journal of AOACInternational,84(3),629-633;以及2009年12月发布的US 7,629,139,如同所充分阐述的,其全部通过引证结合至本文。这些测定通常依赖于将由植酸钠随时间释放的磷酸盐的量与磷酸盐标准曲线进行比较,并调节背景磷酸盐水平和酶水平。测量通常以植酸酶单位(FTU)报告,FTU定义为在给定的一组测定条件下(通常为37℃,pH5.5下,在过量的磷酸钠下,但其他条件也被报道并用在研究和工业中)每单位时间(通常为1分钟)释放的磷酸盐的量(通常为微摩尔的无机磷酸盐)。这些方法可以利用微生物产生的植酸酶和工程化植酸酶,以及从其他宿主表达系统(包括植物表达系统)产生的植酸酶进行。
为了进行测定,必须从表达宿主制备酶提取物。存在多种不同的蛋白质制备方法并且是本领域已知的。在每种方法中,使用诸如机械破碎(例如使用弗氏压碎器)、液体均化、超声、反复冻融循环、去污剂和化学裂解或手动研磨的方法破坏实验细胞(case cell)。细胞裂解后,可以直接使用裂解物,或者可以进一步分馏裂解物以富集所需蛋白质,或者甚至将所述裂解物纯化成几乎纯的蛋白质物质(参见“Current Protocols in MolecularBiology”,10.0.1-10.0。2010年4月23日,John Wiley&Sons,Inc.,如同所充分阐述的,其通过引证结合至本文)。细胞裂解和蛋白提取物甚至可以在很大程度上自动化,便于同时处理许多样品。对于来自植物或种子的蛋白质提取物,通常首先必须破碎较大的组织样品(通常通过碾磨(milling)或研磨(grinding),以及有时包括样品的冷冻或反复的冷冻和解冻循环,并且随后蛋白质可以与以上描述和参考的那些相类似的方法进行提取。
植酸酶活性测量从高达1mL的细胞裂解物开始,将蛋白提取物在测定缓冲液(250mM乙酸钠、pH5.5、1mM氯化钙、0.01%吐温20)中稀释100倍。将七十五(75)微升稀释的提取物或75微升仅为缓冲液的对照分配到圆底96孔板的各孔中。将一百五十微升(150μl)新鲜制备的植酸(9.1mM十二烷基钠盐(dodecasodium salt),来自BiosynthInternational,Staad,瑞士,在测定缓冲液中制备)加入到每个孔中。将平板密封并在37℃温育60分钟。将一百五十(150)微升终止液(20mM钼酸铵、5mM钒酸铵、4%硝酸)加入到每个孔中,通过用吸量管将液体吸移充分混合,并使其在室温下孵育10分钟。将平板在3000×G下离心10分钟,并将100μL澄清的上清液转移至平底96孔板的孔中。将来自每个样品的415nm处的吸光度与阴性对照(仅缓冲液,不含酶)和磷酸钾标准品的吸光度进行比较。通过将50μl磷酸钾标准品(0-1.44mM,在测定缓冲液中制备)与100μl新鲜制备的植酸混合,然后加入100μl终止液来制备标准曲线。
实施例7.测试环化植酸酶的热稳定性
为了确定工程化植酸酶的热稳定性,必须在不同的温度处理后测量工程化植酸酶的活性。可以使用本领域已知的植酸酶测定进行植酸酶活性的测量。植酸酶测定(其可以用于测量植酸酶活性)在本文的实施例6中也进行了描述。尽管可以使用多种不同的方法来研究工程化植酸酶的热稳定性,但是本文使用一种方法作为实施例,同时认识到可以在该分析中使用其他方法、实验设计和测定方法。此外,确切的实验条件可能会根据分析的广度和深度而显著变化。不论用于大规模生产工程化植酸酶的最终生产系统如何,优选的方法使用微生物表达系统来快速生产待测试的工程化植酸酶和可能包括在评估中的其他对照分子。可以用于该评估的微生物表达系统包括大肠杆菌(E.coli)、酿酒酵母(Saccharomycescerevisiae)、毕赤酵母(Pichia pastoris)、芽孢杆菌(Bacillus)、黑曲霉(Aspergillusniger)和里氏木霉(Trichoderma reesei)表达系统,但也可以使用其他系统。在微生物表达系统评估之后,只要这些材料可以使用,重复使用由最终生产系统生产的材料的评估将是有益的。
为了评估工程化植酸酶的热稳定性,期望在不同温度和不同的时间长度,在期望的条件下测试工程化植酸酶和相应的目标植酸酶(没有任意的分子结构与目标植酸酶连接)。理想情况下,这些测试的实验设计将使用已知摩尔量的工程化植酸酶和目标植酸酶,将分子在期望的缓冲液中单独孵育一段时间,范围为零秒(未处理的阴性对照)至30分钟或更长。可以在任意期望的时间间隔进行测量,但如果要在较高温度下测量高于测定背景的活性值,则较短的时间间隔将是必要的。在每次孵育中使用恒定的温度以及缓冲液的pH值。在60℃至高达90℃或更高的范围内的温度对于确定工程化植酸酶相对于其相应目标植酸酶的热稳定性是有意义的。同样地,在2至高达7或更高范围内的pH值对于确定植酸酶在生理学相关的酸度水平下的热稳定性是相关的。孵育后,取出孵育混合物的样品,在标准温度(优选25℃至37℃)和pH(优选5至7)下测量酶活性。然后可以将测量的工程化植酸酶的活性与目标植酸酶进行比较,并且可以确定热稳定性的改善。将目标植酸酶Phy02、Nov9X和CQBscks单独地与本文所述的工程化Phy02植酸酶一起温育。在设定为65℃、70℃、75℃、80℃、85℃和90℃的水浴中,在pH5.5下进行孵育。对于每次孵育,在15秒、40秒、1分钟、1.5分钟、2分钟、3分钟、5分钟、10分钟和15分钟时取出样品。在每次孵育之前,取样品代表零时间点,其中没有发生升高的温度暴露。在零时间点测量的活性在实验中观察到的最大活性的实验变化范围内。从零时间点和每个孵育样品开始,如实施例6中所描述的,活性以一式三份在37℃和pH 5.5下测量。然后将工程化的Phy02植酸酶的活性与目标植酸酶Phy02、Nov9X和CQBscks的活性进行比较。Nov9X在整个处理中表现出最低的活性,Phy02和CQBscks在不同的处理中表现出更高的活性。选择工程化Phy02植酸酶,其在不同处理中相对于目标酶具有增加的活性。
通常时间、实验条件并不太理想,使用对该实施例中描述的方法进行改动的方法。期望的是进行至少一式三份的活性测量以便能够确定在给定的一组条件下活性测量中的变化,但在某些情况下,只有重复或单次测量可能是可行的。在许多情况下,纯化每种工程化酶或目标酶以使用等摩尔浓度是不可行的。通常,考虑到来自给定表达系统的不同植酸酶的表达水平可能相似,这也不是必需的。在这些情况下,孵育中的酶的上样量可以基于培养物体积、裂解物体积、总蛋白质的量或类似的变量。在这些评估中也没有必要使用纯化的酶,因为可以使用热稳定性的相对变化来比较酶并评估热稳定性的改善。为了评估热稳定性的相对变化,通过将在所有随后时间点测量的活性除以在零时间点测量的活性并乘以100,将在给定温度下、随时间点测量的活性水平归一化为零时间点。因此,例如,如果工程化Phy02酶在零时间点被测量为具有1000FTU,并且在给定温度(例如90℃)下进行以下测量:15秒950FTU、40秒902FTU、1分钟857FTU、1.5分钟797FTU、2分钟时741FTU、3分钟669FTU、5分钟时545FTU、10分钟400FTU、15分钟238FTU,则活性测量百分比计算如下:100%(0s)、95%(15s)、90.2%(40s)、85.7%(1m)、79.7%(1.5m)、74.1%(2m)、66.9%(3m)、54.5%(5m)、40.0%(10m)和23.8%(15m)。如果确定目标酶的相应值为100%(0s)、85%(15s)、60.2%(40s)、25.7%(1m)、5.1%(1.5m)、1.3%(2m)1.5%(3m)、0.9%(5m)、0.0%(10m)和0.0%(15m),那么本领域技术人员应该清楚,工程化Phy02植酸酶相对于目标植酸酶具有改善的热稳定性。该方法可以在多个温度和其他pH值下重复以更详细且更精确地限定工程化植酸酶与目标植酸酶之间的热稳定性差异。使用相对测量和容易获得的自动化,可以容易地筛选和评估许多工程化植酸酶变体,并且选择最大改善的酶用于商业用途。
此外,存在其他方法来确定热稳定性。差示扫描量热法是本领域已知的方法,其可以提供非常准确的热稳定性测量。
实施例8.工程化植酸酶的热稳定性优化
前述实施例中描述的任意分子或方法可以在工程化植酸酶的热稳定性或其他特性中继续开发进一步的改善。特定的商业和科学兴趣的特性包括工程化植酸酶的比活性、工程化植酸酶在各种异源表达系统(包括微生物表达系统、植物表达系统和哺乳动物表达系统)中的表达水平、工程化植酸酶的胃和胃蛋白酶稳定性以及工程化植酸酶的胃蛋白酶消化率。存在用于进一步优化工程化植酸酶以具有改善的热稳定性或其他性能的许多方法。这些方法包括定点诱变、饱和诱变、随机诱变、序列洗牌(sequence shuffling)、建模(modeling)等。此外,这些方法可以使用自动筛选系统而容易利用,从而能够在合理的时间范围内评估数百万的变体。
为了优化工程化植酸酶(其编码序列包括内含肽序列),多种方法可以是特别有用的,包括饱和诱变和定点诱变。本领域已知的是,在内含肽-外显肽连接点附近发生的突变可以对内含肽剪接产生显著影响,从而允许这样的分子发展,其能够结合但不剪接,结合并产生异构肽,结合并选择性剪切断裂内含肽的一部分,或者结合并完全剪切以在插入点形成共价键(Xu,MQ,&Perler,F.B.(1996).The mechanism of protein splicing and itsmodulation by mutation.The EMBO journal,15(19),5146,如同所充分阐述的,其通过引证结合至本文)。因此,相对于内含肽插入位点,在内含肽连接处目标植酸酶的-3至-1位置处的突变以及+1至+3位置处的突变,通常对于结合和剪接反应的程度以及不同条件下的反应速率具有显著影响。在这些位点处的突变可以提高剪接速率,从而提高植酸酶的环化速率,以及在某些情况下观察到的酶的热稳定性(如实施例7中所评估的)。因为优选的插入盒(insertion cassette)已经对于许多内含肽进行了鉴定,所以这些盒可以成功用于目标植酸酶骨架中以改善内含肽剪接并因此改善获得的工程化植酸酶的热稳定性,或者用于连接子中以达到相同的目的和效果。类似地,蛋白质编码序列中的其他突变(包括分子结构)可以用于改善热稳定性。对于内含肽的插入盒,参见Apgar等,2012,如同所充分阐述的,该申请通过引证并入本文。
比活性、异源表达水平、胃稳定性和胃蛋白酶消化也可以通过对本研究中构建的工程化植酸酶的进一步的诱变研究而得到改善。用于优化这些性能的方法将以热稳定性优化的类似方式进行,但是在每种情况下,评估程序中会考虑不同的特性。
实施例9.用于工程化植酸酶的表达盒的描述
用于植物表达的环状植酸酶序列和图谱:含有用于植物表达的环状植酸酶的不同变体的序列已经装配成表达盒,在表达盒中KpnI限制性位点在5'处并且EcoRI限制性位点在3'末端。单个遗传元件的所有序列都经过密码子优化以在玉米中表达。每个单独序列的两个盒设计为一个用于胞质靶向蛋白表达,另一个用于内质网(ER)靶向蛋白质表达。为了产生最终的植物表达构建体,可以将每个表达盒克隆到KpnI-EcoRI酶切载体,如pAG4500中。在图8中示出了包括以这样方式克隆的表达盒ZmZ27:Gp41-1C:Phy02opt:Gp41-1N:NosT(Phy02opt盒)的构建体pAG4918的代表性图谱。如图8中所示,包括编码ZmZ27启动子、Gp41-1C内含肽、Phy02opt植酸酶、Gp41-1N内含肽和NosT终止子的多核苷酸的Phy02opt表达盒可以在KpnI位点(位置10227)和EcoR1位点(位置283)引入pAG4918中。pAG4918还携带由玉米泛素(ZmUbi1)启动子、玉米泛素(ZmUbi1)内含子、玉米(Zm)Kozak、磷酸甘露糖异构酶编码序列和NosT终止子、磷酸甘露糖异构酶(PMI)基因和NosT终止子组成的植物选择标签。Phy02opt和植物选择标签盒都在右边界(RT)和左边界(LB)之间整合到pAG4918中。pAG4918包括大观霉素腺苷酰转移酶基因(aadA)、链丝菌素腺苷转移酶基因、噬菌体λ的粘性位点(cos)和复制起点Ori。pAG4918或类似载体可以通过接合转移从大肠杆菌转移到根癌农杆菌LBA4404,在此期间,质粒将通过同源重组整合到pSB1(内生Ti质粒)中。所得到的重组农杆菌菌株与植物细胞的共培养可以导致pAG4918来源的DNA转移至植物基因组。本文的实施方式包括具有任意一种工程化植酸酶的转化载体。
植物转化载体通过将本文所述的表达盒或构建体插入pAG4500或任意合适质粒的农杆菌T-DNA右边界(RB)和左边界(LB)序列之间来组装。
图9A-图9C示出了所选择的工程化植酸酶(具有断裂内含肽连接至植酸酶编码序列的末端)的表达盒的实例。图9A示出了Phy02opt表达盒ZmZ27P:xGZein27ss:Gp41-1C:Phy02opt:Gp41-1N:DPNGSEKDEL:NosT,其包括编码以下各项的多核苷酸:ZmZ27启动子、GZein27ss信号序列、Gp41-1C内含肽、Phy02opt植酸酶、Gp41-1N内含肽、DPNG连接子、SEKDEL末端延伸序列和NosT终止子,它们可以在KpnI位点(位置10227)和EcoR1位点(位置283)引入pAG4918。pAG4918还携带由玉米泛素(ZmUbi1)启动子、玉米泛素(ZmUbi1)内含子、玉米(Zm)Kozak、磷酸甘露糖异构酶(PMI)编码序列和NosT终止子组成的植物选择标签。图9B示出了ZmZ27P:Ssp DnaE-C:Phy02opt:Ssp DnaE-N:NosT表达盒。参考图9B,表达盒包括ZmZ27启动子、Ssp DnaE-C内含肽、Phy02opt植酸酶、Ssp DnaE-N内含肽和NosT终止子。图9C示出了ZmZ27P:xGZein27ss:Ssp DnaE-C:Phy02opt:Ssp DnaE-N:DPNGSEKDEL:NosT表达盒。参考图9C,表达盒包括ZmZ27启动子、GZein27ss信号序列、Ssp DnaE-C内含肽、Phy02opt植酸酶、Ssp DnaE-N内含肽、DPNG连接子、SEKDEL末端延伸序列和NosT终止子。
图10A-10H是示出了所选择的工程化植酸酶的表达盒的示意图,所述工程化植酸酶具有与连接至植酸酶编码序列末端的连接子连接的断裂内含肽。
图10A示出了ZmZ27P:Ssp DnaE-C:L33-1:Phy02opt:L33-2:Ssp DnaE-N:NosT表达盒。参考图10A,表达盒包括ZmZ27启动子、Ssp DnaE-C内含肽、L33-1连接子(L33-1)、Phy02opt植酸酶、L33-2连接子(L33-2)、Ssp DnaE-N内含肽和NosT终止子。图10B示出了ZmZ27P:xGZein27ss:Ssp DnaE-C:L33-1:Phy02opt:L33-2:Ssp DnaE-N:DPNGSEKDEL:NosT表达盒。参考图10B,表达盒包括ZmZ27启动子、GZein27ss信号序列、Ssp DnaE-C内含肽、L33-1连接子(L33-1)、Phy02opt植酸酶、L33-2连接子(L33-2)、Ssp DnaE-N内含肽、DPNG连接子、SEKDEL末端延伸序列和NosT终止子。图10C示出了ZmZ27P:Ssp DnaE-C:L38-1:Phy02opt:L38-2:Ssp DnaE-N:NosT表达盒。参考图10C,表达盒包括ZmZ27启动子、SspDnaE-C内含肽、L38-1连接子(L38-1)、Phy02opt植酸酶、L38-2连接子(L38-2)、Ssp DnaE-N内含肽和NosT终止子。图10D示出了ZmZ27P:xGZein27ss:Ssp DnaE-C:L38-1:Phy02opt:L38-2:Ssp DnaE-N:DPNGSEKDEL:NosT表达盒。参考图10D,表达盒包括ZmZ27启动子、GZein27ss信号序列、Ssp DnaE-C内含肽、L38-1连接子(L38-1)、Phy02opt植酸酶、L38-2连接子(L38-2)、Ssp DnaE-N内含肽、DPNG连接子、SEKDEL末端延伸序列和NosT终止子。图10E示出了ZmZ27P:Ssp DnaE-C:L46-1:Phy02opt:L46-2:Ssp DnaE-N:NosT表达盒。参考图10E,表达盒包括ZmZ27启动子、Ssp DnaE-C内含肽、L46-1连接子(L46-1)、Phy02opt植酸酶、L46-2连接子(L46-2)、Ssp DnaE-N内含肽和NosT终止子。图10F示出了ZmZ27P:xGZein27ss:SspDnaE-C:L46-1:Phy02opt:L46-2:Ssp DnaE-N:DPNGSEKDEL:NosT表达盒。参考图10F,表达盒包括ZmZ27启动子、GZein27ss信号序列、Ssp DnaE-C内含肽、L46-1连接子(L46-1)、Phy02opt植酸酶、L46-2连接子(L46-2)、Ssp DnaE-N内含肽、DPNG连接子、SEKDEL末端延伸序列和NosT终止子。图10G示出了ZmZ27P:Ssp DnaE-C:L55-1:Phy02opt:L55-2:Ssp DnaE-N:NosT表达盒。参考图10G,表达盒包括ZmZ27启动子、GZein27ss信号序列、Ssp DnaE-C内含肽、L55-1连接子(L55-1)、Phy02opt植酸酶、L55-2连接子(L55-2)、Ssp DnaE-N内含肽、DPNG连接子、SEKDEL末端延伸序列和NosT终止子。图10H示出了ZmZ27P:xGZein27ss:Ssp DnaE-C:L55-1:Phy02opt:L55-2:Ssp DnaE-N:DPNGSEKDEL:NosT表达盒。参考图10H,表达盒包括ZmZ27启动子、GZein27ss信号序列、Ssp DnaE-C内含肽、L55-1连接子(L55-1)、Phy02opt植酸酶、L55-2连接子(L55-2)、Ssp DnaE-N内含肽、DPNG连接子、SEKDEL末端延伸序列和NosT终止子。图10A-图10H中示出的每个盒子具有KpnI、EcoRI和BamHI限制性位点,并且可以作为KpnI-EcoRI片段克隆到转化载体的T-DNA中。
表6中汇编了表达构造体的列表。
表6构造体列表
除了两个连接子序列以外,载体pAG4924、pAG4926和pAG4928中的核苷酸序列与pAG4922中的相同。类似地,除两个连接子序列外,构建体pAG4925、pAG4927和pAG4929中的所有核苷酸序列与pAG4923中的相同。在提供的表达盒pAG4918-pAG4929的图谱上指定的连接子序列包括L33-1、L33-2、L38-1、L38-2、L46-1、L46-2、L55-1和L55-2,并且示出了在表4中。
用于环状植酸酶的植物表达盒的相关序列
>ZmZ27P:Gp411C:Phy02opt:Gp411N:NoST
ZmZ27P以粗体大写字体和斜体示出,gp411用下划线表示,NosT用斜体表示。
>ZmZ27P:xGZein27ss:Gp411-C:Phy02opt:Gp411-N:DPNGSEKDEL:NOST
ZmZ27P以粗体大写字体和斜体显示,gp411以下划线显示,DPNG以大写斜体显示,SEKDEL以粗体大写显示,NosT以斜体显示。
>ZmZ27P:Ssp_DnaE-C:Phy02opt:Ssp_DnaE-N:NOsT
ZmZ27P以粗体大写字体并用斜体表示,SSp_DnaE用下划线表示,NosT用斜体表示。
>ZmZ27P:xGZein27ss:Ssp_DnaE-C:Phy02opt:Ssp_DnaE-N:DPNGSEKDEL:NOsT
ZmZ27P以粗体大写字体和斜体显示,Ssp_DnaE以下划线显示,DPNG以大写斜体显示,SEKDEL以粗体大写显示,NosT以斜体显示。
>ZmZ27P:Ssp_DnaE:L33-1:Phy02opt:L33-3:NOST(SSp_DnaE-C:L33-1:Phy02opt:L33-2:Ssp_DnaE-N)
ZmZ27P以粗体大写字体和斜体表示,Ssp_DnaE以下划线表示,连接子以粗体表示,DPNG以大写斜体表示,SEKDEL以粗体大写表示,以及NosT以斜体表示。
>ZmZ27P:xGZein27ss:Ssp_DnaE:L33-1:Phy02opt:L33-2:DPNGSEKDEL:NOsT
ZmZ27P用粗体大写字体和斜体表示,Ssp_DnaE用下划线表示,L33连接子用粗体大写字母表示,DPNG用大写字母斜体表示,SEKDEL用粗体大写字母表示,以及NosT用斜体表示。
实施例11环化植酸酶在转基因植物中的表达
将如上所述的用载体转化的独立的转基因玉米植物生长至成熟,并用野生型(未转化的)玉米植物进行交叉授粉。从这些植物的每个中收获大约20颗种子。种子通过0.5毫米的筛子研磨以产生细粉末。然后提取酶并如以下所述的测定植酸酶活性。
来自种子的植酸酶测定,方案的简要说明:通过将15mg研磨的种子粉在室温下,在1.5ml的25mM的硼酸钠,pH10,0.01%吐温20中温育1小时来制备酶提取物。然后将提取物在测试缓冲液(250mM乙酸钠,pH5.5,1mM氯化钙,0.01%吐温20)中进行100倍稀释,将七十五(75)微升稀释的提取物或75微升仅为缓冲液的对照物分配到圆底96孔板的各个孔中。将一百五十(150)微升新鲜制备的植酸(9.1mM十二烷基钠盐,来自Biosynth International,Staad,瑞士,在测试缓冲液中制备)加入到每个孔中。将平板密封并在37℃温育60分钟。向每个孔中加入150μL终止液(20mM的钼酸铵、5mM的钒酸铵、4%硝酸),通过用吸量管将液体吸移充分混合,并允许在室温下孵育10分钟。将平板在3000×G下离心10分钟,并将100μL澄清的上清液转移至平底96孔板的孔中。将每个样品在415nm处的吸光度与阴性对照(仅缓冲液,不含酶)和磷酸钾标准品的吸光度进行比较。通过将50μl的磷酸钾标准品(0-1.44mM,在测试缓冲液中制备)与100μl新鲜制备的植酸混合,然后加入100μl的终止液来制备标准曲线。
如所预期的,来独立的转基因植物的种子中的植酸酶活性变化显著。
实施例12.造粒过程中环状植酸酶的热稳定性
为了确定工程化植酸酶的热稳定性,饲料必须混合为含有特定水平的工程化植酸酶、相应的目标植酸酶和任意对照植酸酶,期望比较热稳定性,并且包括在评价中。为了测试饲料的热稳定性,在一些不同剂量水平下混合多种日粮是有益的,然后在不同温度下进行一系列造粒过程来评估每种日粮。评估中使用的剂量可以包括500FTU/kg、1000FTU/kg或3000FTU/kg。评估中使用的温度可以包括60℃、65℃、70℃、75℃、80℃、85℃、90℃和95℃或任意其他期望的温度。造粒过程中的停留时间范围可以为15秒或更短至1分钟或更长。对于每种配制的日粮,对于每种酶(和不含酶的阴性对照日粮),除了在造粒之后采集的样品之外,还采集预造粒样品。由测量并比较这些样本的活性。在每次处理中,将颗粒样品与相应的糊状样品(mash sample)进行比较,并与使用包括在实验中的其他酶进行的相同处理进行比较。造粒后,在最高温度下保持最高活性百分比的工程化酶,显示出了最大程度的热稳定性。比相应目标植酸酶,表现出更高热稳定性的工程化植酸酶具有改善的热性能,并且是用于商业开发的候选者。
实施例13.环状植酸酶在肉鸡和猪中的性能
用低含量的无机磷酸盐制备玉米-大豆基础日粮。由这些基础日粮,通过添加量子蓝(Quantum Blue)(AB酶)形式的酶或表达Phy02、Nov 9X、工程化环状Phy02或工程化环状Nov9X的碾磨玉米籽粒,并变化掺入杯中日粮中的酶的总量,来制备重复日粮。对于Phy02和Nov9X,从基础日粮中除去少量玉米来说明转基因谷物被添加了回来以供应酶。制备对照日粮,其中相对于基础日粮,无机磷酸盐的量增加。
雄性肉仔鸡在栏中被分配在不同的饲料处理中,每栏约12只鸡,并且每个处理6个重复栏。将饲料以糊状(mash)形式提供给一组鸡,并且将颗粒饲料提供给另一组鸡。在14、21、28、35和42天后,对鸡进行称重并比较以确定各种酶处理对肉鸡生产的影响。
类似地,将猪在栏中被分配在不同的饲料处理中,每栏约7只猪,每个处理5个重复栏。为猪提供颗粒饲料。在21、35和49天后,对猪进行称重并比较以确定各种酶处理对猪生产的影响。
实施例14修饰的植酸酶的热稳定性
图11示出了SspDnaE-C:Phy02:SspDnaE-N构建体的表达图谱。参考该图,“C”表示粗提取物,“S”表示可溶部分,“*”标志了粗提取物中目标蛋白质的位置,“○”标示了粗提取物中环状Phy02的位置。IPTG诱导的表达培养物的考马斯凝胶。将构建体克隆在pETDuetI(Novagen)的EcoRI和XhoI位点之间并转化到Shuffle T7(NEB)大肠杆菌表达宿主中。为了分析表达图谱,将LB+羧苄青霉素(100mg/L)中的过夜的发酵培养物以40倍稀释至新鲜培养基并在30℃,250rpm下生长至OD600=0.6,然后加入IPTG至终浓度为0.5mM,培养物再生长另外3个小时。以3000g离心10分钟收获细胞,用一倍培养体积的植酸酶洗涤缓冲液(250mMNaOAc pH=5.5和1mM CaCl2)洗涤,并如前所述将细胞沉淀。将细胞沉淀物在含有1XFastbreak(Promega)(含有核酸酶(Benzonase)(50U/mL,Novagen))的植酸酶裂解缓冲液中裂解(30℃,250rpm,1hr)。用于考马斯凝胶的样品制备如下:通过将等体积的裂解物与含有5%β-巯基乙醇的2X Laemmli样品缓冲液(Bio-Rad)混合来制备粗提物(C)。为了制备可溶部分(S),裂解物以5000g离心10分钟,如前所述将上清液与等体积的上样染料混合。热可溶部分(H)通过在55℃孵育裂解物15分钟,然后5000g离心10分钟,并将上清液与等体积的上样染料混合来制备。上样前,SDS/PAGE样品在95℃加热5分钟,连同10μLMW标记物(Precision Plus Protein Kaleidoscope,Bio-Rad),将5μL等分试样上样至标准XT 12%Bis-Tris凝胶。分离蛋白质后,用SimplyBlue Safe Stain(Life Technologies的Novex)染色凝胶。
参考图11,观察到Phy02在粗提物(C)、可溶部分(S)和热可溶部分(H)表现相当。SspDnaE-C:phy02:SspDnaE-N融合蛋白的表达水平是可比较的,但在溶解度上表现出显著的差异:没有连接子(-),蛋白质基本是不可溶的,而含有连接子的构建体主要表达可溶性部分,并且也在热可溶部分中得到了很好地显示。除了具有最长连接子(连接子46和55)的两种构建体外,即除了线性蛋白质在粗提物(C)、可溶部分(S)和热可溶部分(H)中以相当的水平表现出了更快移动的新蛋白质物种(标记为“o”)之外,Phy02及其内含肽修饰形式在线性分子的期望大小(标签为“*”,大约58KD)处溶解(resolved)。通过比较环化组分SpyTag:Phy02:SpyCatcher与环化缺陷突变体的迁移率(参见图12),确定了较高的迁移率是环状Phy02的标志。
图12示出了Phy02的热稳定性测定。参考图12,粗提取物如图11所述方法制备,并在植酸酶洗涤缓冲液中稀释50倍。将PCR管中的150μL等分试样在PCR模块(block)中热处理,所述PCR模块被编程为用于相同的模块盖温度。在指定的时间点取出管并在室温下孵育1小时以允许重新折叠。将每个样品稀释至250-、1000-、5000-和20000-倍,并基于已建立的方案测定植酸酶活性。
该图示出了在30秒间隔内取样的样品中,在70℃、75℃、80℃和85℃预处理4分钟,粗制细胞裂解物中未修饰的Phy02的热稳定性。仅在70℃/30秒样品中保留了完全的活性。增加热暴露时间和/或温度会迅速降低植酸酶活性。暴露于75℃或80℃下1分钟后,未经修饰的Phy02植酸酶活性分别降至临界可检测或不可检测的水平。
图13A-13B示出了SspDnaE-C:Phy02:SspDnaE-N构建体的热稳定性。表达培养物以及制备粗提取物如图11所示。热预处理在75℃下进行60秒,并如图12所示的测定植酸酶活性。图13A示出了未处理(37℃)和热处理(75℃/60秒)样品的酶活性。图13B示出了热预处理样品中残留的植酸酶活性,以其各自未处理对照(37℃)的活性的百分比表示。
每个连接子修饰的反式-剪接Phy02在热预处理后保留一些活性,该热预处理完全消除未修饰的Phy02对照的植酸酶活性。具有最长连接子(连接子46和55)的两个克隆在热预处理样品中表现出最高的耐热性,保留了约10%的活性。不含连接子的内含肽融合(DnaE-sPhy02_DnaE)并未改善热稳定性。
图14示出了SpyTag:Phy02:SpyCatcher野生型和突变型的表达图谱。IPTG诱导的表达培养物的考马斯凝胶。
将构建体克隆在pETDuetI(Novagen)的NcoI和XhoI位点之间,并转化至ShuffleT7(NEB)大肠杆菌表达宿主中。环化缺陷型突变体在SpyTag中携带丙氨酸突变(野生型AHIVMVDAYKPTK[SEQ ID NO:216]和突变型AHIVMVAAYKPTK[SEQ ID NO:217])。诱导培养物、制备的粗提物(C)、可溶部分(S)和热可溶部分(H)以及SDS/PAGE与图11中的相同。粗提物中的目标蛋白的位置用星号表示。
野生型和突变的SpyTag:Phy02:SpyCatcher均表达可溶部分,并在热可溶部分(H)和可溶部分(S)以及粗提物(C)中表示相同。尽管环化竞争形式(wt)在63kD(552个氨基酸)的预期大小处分离了线性分子,但环化缺陷突变体(mut)在凝胶上快速移动。该观察与SpyTag和SpyCatcher之间的分子内相互作用导致环化竞争分子的分子内环化的解释是一致的。SpyTag中的突变阻止了环化。环状Phy02比环化缺陷的线性分子具有更高的迁移率。环化竞争野生型SpyTag:Phy02:SpyCatcher主要表达高迁移率的Phy02形式,表明环化是高效率的。
图15A表明SpyTag:Phy02:SpyCatcher改善了植酸酶的耐热性,热预处理样品的植酸酶活性。重组蛋白的表达如图14中所示,热预处理和酶测定如图12所述在75℃和80℃中进行,并且等分试样在120秒内以30秒间隔取样。分别地,左部分(panel)示出了在75℃热处理后的酶活性,右部分示出了在80℃热处理后的酶活性。环化竞争野生型SpyTag:Phy02:SpyCatcher(wt)示出了出显著改善的热稳定性,并且在80℃,在测试的整个热预处理的长度上保持稳定。与未修饰的Phy02相比,环化缺陷型突变体SpyTag:Phy02:SpyCatcher(mut)也表现出改善的热稳定性。
图15B示出了SpyTag:Phy02:SpyCatcher提高了植酸酶的耐热性,热预处理样品的植酸酶活性的保留。图17A的热预处理样品的植酸酶活性以它们各自的未处理对照的百分比表示进行绘制。环化竞争植酸酶(wt)在80℃保持超过35%的活性,在整个2分钟的热处理期间保持稳定。相反,环化无效线性形式(mut)在80℃迅速失去活性,但耐热性超过未修饰的Phy02的热稳定性。这种有益效果可能是由于在环化无效突变SpyTag构建体中SpyCatcher的重折叠功能的保留。可能地,环状和线性分子的植酸酶活性耐热性之间的差异可以表明环化和再折叠对热稳定性的影响程度。
实施例15.需要内含肽剪接以获得环状植酸酶构建体的高耐热性
通过使用刚性连接子55-1和55-2以及反式剪接内含肽gp41-1构建原型环状植酸酶,创建了gp41-1C:L55-1:Phy02:L55-2:gp41-1N[氨基酸(AA)_SEQ ID NO:201和核酸(NA)_SEQ ID NO:200]。此外,创建了构建体的溶解优化的形式,其在N-末端处具有溶解性增强剂硫氧还蛋白结构域(TrxH)[AA_SEQ ID NO:197和NA_SEQ ID NO:196],该N-末端处,连接有Asp-Pro-Asn-Gly连接子(DPNG)[AA_SEQ ID NO:199和NA_SEQ ID NO:198]至编码TrxH:DPNG:gp41-1C[MTT]:L55-1:Phy02:L55-2:gp41-1N[AA_205和NA_204]的gp41-1C[MTT]的突变形式。
将构建体克隆于来自Shuffle T7大肠杆菌宿主的pETDuetI的EcoRI和XhoI位点之间,并测试植酸酶热稳定性。诱导培养物和粗裂解物的制备如图11所述。对于热处理,将PCR管中的150μL粗裂解物在特定温度下在PCR模块中加热1分钟,然后将该管在室温下孵育1小时以允许再折叠。将每个样品稀释至250倍、1000倍、5000倍和2000倍,并且如图12所示测定植酸酶活性。图16示出了,与野生型酶Phy02(垂直线)和空载体(水平线)相比,环状植酸酶gp41-L55-1:Phy02:L55-2:gp41-1N(闭合圆)和TrxH:DPNG:gp41-1C[MTT]:L55-1:Phy02:L55-2:gp41-1N(闭合正方形)的热预处理。参考图16,观察到野生型植酸酶(Phy02)在75℃以上迅速丧失活性,而两种环状植酸酶构建体在85℃保持活性,在原型对溶解度优化的植酸酶中分别显示了16%对8%的活性。
为了评估是否需要蛋白环化来获得耐热性,通过在以下两种环状植酸酶构建体中,利用不同的连接子来突变剪接必需的氨基酸残基使得剪接失效:TrxH:DPNG:gp41-1C[MTT]:L46-:1C Phy02:L46-2:gp41-1N[AA_SEQ ID NO:207和NA_SEQ ID NO:206]和gp41-1C[MTT]:L55-1:Phy02:L55-2:gp41-1N[AA_SEQ ID NO:205和NA_SEQ ID NO:204]。剪接失效突变是gp41-1C内含肽C-末端Asn残基变为Ala残基[N125A],或连接子+1位上,gp41-1CC-末端侧翼Ser残基变为Ala残基[S1A]。创建了下列突变体:[N125A-1]剪接无效TrxH:DPNG:gp41-1C[MTT]:L46-1:Phy02:L46-2:gp41-1N[AA_SEQ ID NO:209和NA_SEQ ID NO:208]、[N125A-2]剪接无效gp41-1C[MTT]:L55-1:Phy02:L55-2:gp41-1N[AA_SEQ ID NO:213和NA_SEQ ID NO:212]、[S1A-1]剪接无效TrxH:DPNG:gp41-1C[MTT]:L46-1:Phy02:L46-2:gp41-1N[AA_SEQ ID NO:211和NA_SEQ ID NO:210]和[S1A-2]剪接无效gp41-1C[MTT]:L55-1:Phy02:L55-2:gp41-1N[AA_SEQ ID NO:215和NA_SEQ ID NO:214]。
在Shuffle T7大肠杆菌宿主中表达的pETDuetI的EcoRI和XhoI位点之间克隆构建体,并且在85℃/1分钟的热预处理之后测试剪接有效和无效的建体提的耐热性。图17示出了在85℃预处理1分钟后,剪接有效的和剪接无效的(内含肽N125A和连接子S1A)环状植酸酶gp41-1C:L55-1:Phy02:L55-2:gp41-1N和TrxH:DPNG:gp41-1C[MTT]:L55-1:Phy02:L55-2:gp41-1N和野生型Phy02植酸酶的植酸酶活性。参考图17,观察到在37℃,所有构建体表现出植酸酶活性。然而,在暴露于85℃下1分钟后,只有剪接有效的构建体保持活性。剪接无活的突变体都表现出与内含肽未修饰的野生型植酸酶相似的热敏性。这些结果与获得耐热性取决于内含肽剪接介导的蛋白质环化的解释是一致的。
参考文献
Apgar,J.,Ross,M.,Zuo,X.,Dohle,S.,Sturtevant,D.,Shen,B.,...&Raab,R.M.(2012).A predictive model of intein insertion site for use in the engineeringof molecular switches.PloS one,7(5),e37355.
Arakawa,T.,Chong,D.K.,&Langridge,W.H.(1998).Efficacy of a food plant-based oral cholera toxin B subunit vaccine.Nature Biotechnology,16(3),292–297.doi:10.1038/nbt0398-292.
Basu,S.S.,Winslow,S.,Nelson,A.,Ono,M.,&Betts,S.(2009).EXTRACTIONMETHODS AND ASSAYS FOR FEED ENZYMES.
Cervelli,M.,Di Caro,O.,Di Penta,A.,Angelini,R.,Federico,R.,Vitale,A.,&Mariottini,P.(2004).A novel C-terminal sequence from barley polyamineoxidase is a vacuolar sorting signal.Plant Journal,40(3),410–418.doi:10.1111/j.1365-313X.2004.02221.X.
“Current Protocols in Molecular Biology,”10.0.1-10.0.23,April,2010,John Wiley&Sons,Inc.
Engelen,A.J.,Heeft,F.C.,Randsdorp,P.H.,Somers,W.A.,Schaefer,J.,&vander Vat,B.J.(2001).Determination of phytase activity in feed by acolorimetric enzymatic method:collaborative interlaboratory study.Journal ofAOAC International,84(3),629-633.
FU,T.J.(2002).Digestion stability as a criterion for proteinallergenicity assessment.Annals of the New York Academy of Sciences,964(1),99-110.
Gogarten,J.P.,Senejani,A.G.,Zhaxybayeva,O.,Olendzenski,L.,&Hilario,E.(2002).Inteins:structure,function,and evolution.Annual Reviews inMicrobiology,56(1),263-287.
Haq,T.a,Mason,H.S.,Clements,J.D.,&Arntzen,C.J.(1995).Oralimmunization with a recombinant bacterial antigen produced in transgenicplants.Science(New York,N.Y.),268(5211),714–716.doi:10.1126/science.7732379.
Lau SYM,Taneja AK and Hodges RS(1984)Synthesis of a model protein ofdefined secondary and quaternary structure.Effect of chain length on thestabilization and formation of two-strandedα-helical coiled-coils.J.Biol.Chem.259(21),13253-61.
Korban,S.S.(2002).Targeting and expression of antigenic proteins intransgenic plants for production of edible oral vaccines.In Vitro Cellular&Developmental Biology-Plant,38(3),231–236.doi:10.1079/IVP2002292.
Munro,S.,&Pelham,H.R.(1987).A C-terminal signal prevents secretion ofluminal ER proteins.Cell,48(5),899–907.doi:10.1016/0092-8674(87)90086-9.
Parry DA,Fraser RD and Squire JM(2008)Fifty years of coiled-coils andalpha-helical bundles:a close relationship between sequence and structure.JStruct Biol.163(3),258-69.
Perler,F.B.(2002).InBase:the intein database.Nucleic acids research,30(1),383-384.
Perler,F.B.,Davis,E.O.,Dean,G.E.,Gimble,F.S.,Jack,W.E.,Neff,N.,...&Belfort,M.(1994).Protein splicing elements:inteins and exteins--a definitionof terms and recommended nomenclature.Nucleic acids research,22(7),1125.
Schoene,C.,Fierer,J.O.,Bennett,S.P.,&Howarth,M.(2014).SpyTag/SpyCatcher cyclization confers resilience to boiling on a mesophilicenzyme.Angewandte Chemie International Edition,53(24),6101-6104.
Thomas,K.,Aalbers,M.,Bannon,G.A.,Bartels,M.,Dearman,R.J.,Esdaile,D.J.,...&Zawodny,J.(2004).A multi-laboratory evaluation of a common in vitropepsin digestion assay protocol used in assessing the safety of novelproteins.Regulatory Toxicology and Pharmacology,39(2),87-98.
US 7,629,139,issued December 8,2009.
Woolfson DN(2005)The design of coiled-coil structures andassemblies.Adv.Protein Chem.70,79-f112
Xu,M.Q.,&Perler,F.B.(1996).The mechanism of protein splicing and itsmodulation by mutation.The EMBO journal,15(19),5146.
Zakeri,B.,Fierer,J.O.,Celik,E.,Chittock,E.C.,Schwarz-Linek,U.,Moy,V.T.,&Howarth,M.(2012).Peptide tag forming a rapid covalent bond to aprotein,through engineering a bacterial adhesin.Proceedings of the NationalAcademy of Sciences,109(12),E690-E697.
如同每个参考文献所充分阐述的,整个本申请中引用的参考文献结合用于本文以及参考文献本身中显而易见的所有目的。为了呈现,这些参考文献中的特定文献在本文中的特定位置被引用。在特定位置引用参考文献表示其中教导的参考文献结合的方式。然而,在特定地点引用参考文献并不限制教导的引用参考文献结合用于所有目的的方式。
因此,应该理解的是,本发明不限于所公开的特定实施方式,而是旨在覆盖由所附权利要求限定的在本发明的精神和范围内的所有修改;以上描述;和/或在附图中所示出的。
序列表
<110> 谷万达公司
<120> 工程化植酸酶及其使用方法
<130> P50142VOL
<150> US62/220,688
<151> 2015-09-18
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Leu Ala Asn Gly Ile Phe Val His Asn
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Asn Gly Ile Leu Val His Asn
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Lys Gly Asp Arg Ile Ile Val Ala Lys
115 120
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<212> PRT
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Met Thr His Val Leu Phe Asp Glu Ile Val Glu Ile Arg Tyr Ile Ser
1 5 10 15
Glu Gly Gln Glu Val Tyr Asp Val Thr Thr Glu Thr His Asn Phe Ile
20 25 30
Gly Gly Asn Met Pro Thr Leu Leu His Asn
35 40
<210> 17
<211> 111
<212> PRT
<213> Artificial Sequence
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Cys Leu Leu Pro Glu Glu Arg Val Ile Leu Pro Asp Tyr Gly Pro Ile
1 5 10 15
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20 25 30
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35 40 45
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50 55 60
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85 90 95
Arg Ala Asp Gln Leu Lys Pro Gly Asp Tyr Val Ala Val Pro Lys
100 105 110
<210> 18
<211> 39
<212> PRT
<213> Artificial Sequence
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Met Asn Phe Val Phe Leu Pro Val Glu Lys Ile Glu Glu Phe Glu Tyr
1 5 10 15
Asp Gly Tyr Val Tyr Asp Val Thr Thr Glu Thr His Asn Phe Ile Ala
20 25 30
Asn Gly Ile Leu Val His Asn
35
<210> 19
<211> 121
<212> PRT
<213> Artificial Sequence
<400> 19
Cys Val Asp Gly Asp Thr Leu Val Leu Thr Lys Glu Phe Gly Leu Ile
1 5 10 15
Lys Ile Lys Glu Leu Tyr Glu Lys Leu Asp Gly Lys Gly Arg Lys Ile
20 25 30
Val Glu Gly Asn Glu Glu Trp Thr Glu Leu Glu Lys Pro Ile Thr Val
35 40 45
Tyr Gly Tyr Lys Asp Gly Lys Ile Val Glu Ile Lys Ala Thr His Val
50 55 60
Tyr Lys Gly Val Ser Ser Gly Met Val Glu Ile Arg Thr Arg Thr Gly
65 70 75 80
Arg Lys Ile Lys Val Thr Pro Ile His Arg Leu Phe Thr Gly Arg Val
85 90 95
Thr Lys Asp Gly Leu Ile Leu Lys Glu Val Met Ala Met His Val Lys
100 105 110
Pro Gly Asp Arg Ile Ala Val Val Lys
115 120
<210> 20
<211> 42
<212> PRT
<213> Artificial Sequence
<400> 20
Met Gln His Ile Ile Phe Asp Glu Val Ile Asp Val Arg Tyr Ile Pro
1 5 10 15
Glu Pro Gln Glu Val Tyr Asp Val Thr Thr Glu Thr His Asn Phe Val
20 25 30
Gly Gly Asn Met Pro Thr Leu Leu His Asn
35 40
<210> 21
<211> 96
<212> PRT
<213> Artificial Sequence
<400> 21
Cys Leu Ala Gly Asp Thr Leu Ile Thr Leu Ala Asp Gly Arg Arg Val
1 5 10 15
Pro Ile Arg Glu Leu Val Ser Gln Gln Asn Phe Ser Val Trp Ala Leu
20 25 30
Asn Pro Gln Thr Tyr Arg Leu Glu Arg Ala Arg Val Ser Arg Ala Phe
35 40 45
Cys Thr Gly Ile Lys Pro Val Tyr Arg Leu Thr Thr Arg Leu Gly Arg
50 55 60
Ser Ile Arg Ala Thr Ala Asn His Arg Phe Leu Thr Pro Gln Gly Trp
65 70 75 80
Lys Arg Val Asp Glu Leu Gln Pro Gly Asp Tyr Leu Ala Leu Pro Arg
85 90 95
<210> 22
<211> 40
<212> PRT
<213> Artificial Sequence
<400> 22
Met Ser Asp Val Tyr Trp Asp Pro Ile Val Ser Ile Glu Pro Asp Gly
1 5 10 15
Val Glu Glu Val Phe Asp Leu Thr Val Pro Gly Pro His Asn Phe Val
20 25 30
Ala Asn Asp Ile Ile Ala His Asn
35 40
<210> 23
<211> 89
<212> PRT
<213> Artificial Sequence
<400> 23
Cys Leu Gly Lys Gly Thr Pro Val Met Met Tyr Asp Gly Arg Thr Lys
1 5 10 15
Pro Val Glu Lys Val Glu Val Gly Asp Arg Leu Met Gly Asp Asp Gly
20 25 30
Ser Pro Arg Thr Val Gln Ser Leu Ala Arg Gly Arg Glu Gln Met Tyr
35 40 45
Trp Val Arg Gln Lys Arg Gly Met Asp Tyr Arg Val Asn Glu Ser His
50 55 60
Ile Leu Ser Leu Lys Lys Ser Arg Arg Glu Gly Ala Arg Asp Arg Gly
65 70 75 80
Ser Ile Ala Asp Ile Ser Val Arg Asp
85
<210> 24
<211> 38
<212> PRT
<213> Artificial Sequence
<400> 24
Met Trp Arg Met Thr Gly Ile Asp Val Glu Pro Asp Gly Val Gly Asp
1 5 10 15
Tyr Phe Gly Phe Thr Leu Asp Gly Asn Gly Arg Phe Leu Leu Gly Asp
20 25 30
Gly Thr Val Thr His Asn
35
<210> 25
<211> 95
<212> PRT
<213> Artificial Sequence
<400> 25
Cys His Pro Ala Asp Thr Lys Val Ile Val Lys Gly Lys Gly Ile Val
1 5 10 15
Asn Ile Ser Asp Val Lys Glu Gly Asp Tyr Ile Leu Gly Ile Asp Gly
20 25 30
Trp Gln Arg Val Lys Lys Val Trp Lys Tyr His Tyr Glu Gly Lys Leu
35 40 45
Ile Asn Ile Asn Gly Leu Lys Cys Thr Pro Asn His Lys Val Pro Val
50 55 60
Val Thr Glu Asn Asp Arg Gln Thr Arg Ile Arg Asp Ser Leu Ala Lys
65 70 75 80
Ser Phe Leu Ser Gly Lys Val Lys Gly Lys Ile Ile Thr Thr Lys
85 90 95
<210> 26
<211> 41
<212> PRT
<213> Artificial Sequence
<400> 26
Met Asn Ser Phe Tyr Asn Leu Ser Thr Phe Glu Val Ser Ser Glu Tyr
1 5 10 15
Tyr Lys Gly Glu Val Tyr Asp Leu Thr Leu Glu Gly Asn Pro Tyr Tyr
20 25 30
Phe Ala Asn Gly Ile Leu Thr His Asn
35 40
<210> 27
<211> 107
<212> PRT
<213> Artificial Sequence
<400> 27
Cys Leu Asp Lys Thr Ala Leu Arg Ile Phe Asn Gln Gly Leu Leu Tyr
1 5 10 15
Ala Asp Glu Val Val Thr Pro Gly Ser Gly Glu Thr Val Gly Leu Gly
20 25 30
Leu Thr Val Arg Asn Gly Ile Gly Ala Ser Thr Ala Ile Ala Asn Gln
35 40 45
Pro Met Glu Leu Val Glu Ile Lys Leu Ala Asn Gly Arg Lys Leu Arg
50 55 60
Met Thr Pro Asn His Arg Met Ser Val Lys Gly Lys Trp Ile His Ala
65 70 75 80
Cys Asn Leu Lys Pro Gly Met Leu Leu Asp Tyr Ser Ile Gly Glu Tyr
85 90 95
Gln Lys Arg Glu Asp Thr Leu Leu Ile Pro Leu
100 105
<210> 28
<211> 49
<212> PRT
<213> Artificial Sequence
<400> 28
Met Ser Lys Cys Val Leu Asn Tyr Ser Pro Tyr Lys Ile Glu Ser Val
1 5 10 15
Asn Ile Gly Ala Val Cys Asp Tyr Ser Tyr Asp Phe Ala Ile Glu Gly
20 25 30
Ile Asn Asp Asn Asp Ser Trp Tyr Trp Gln Gly Ala Leu Lys Ser His
35 40 45
Asn
<210> 29
<211> 111
<212> PRT
<213> Artificial Sequence
<400> 29
Cys Leu Leu Pro Asp Glu Lys Val Ile Leu Pro Glu His Gly Pro Ile
1 5 10 15
Thr Leu Lys Gly Leu Phe Asp Leu Ala Lys Glu Thr Val Val Ala Asp
20 25 30
Asn Glu Lys Glu Ile Arg Lys Leu Gly Ala Lys Leu Thr Ile Val Gly
35 40 45
Glu Asp Gly Arg Leu Arg Val Leu Glu Ser Pro Tyr Val Trp Lys Val
50 55 60
Arg His Arg Gly Lys Met Leu Arg Val Lys Leu Lys Asn Trp His Ser
65 70 75 80
Val Ser Val Thr Pro Glu His Pro Phe Leu Thr Thr Arg Gly Trp Val
85 90 95
Arg Ala Asp Gln Leu Lys Pro Gly Asp Tyr Val Ala Val Pro Arg
100 105 110
<210> 30
<211> 39
<212> PRT
<213> Artificial Sequence
<400> 30
Met Asn Leu Val Phe Ile Pro Val Glu Asp Ile Glu Glu Phe Glu Tyr
1 5 10 15
Glu Gly Tyr Val Tyr Asp Val Thr Thr Glu Thr His Asn Phe Val Ala
20 25 30
Asn Gly Ile Leu Val His Asn
35
<210> 31
<211> 99
<212> PRT
<213> Artificial Sequence
<400> 31
Cys Leu Pro Ala Arg Ala Arg Val Val Asp Trp Cys Thr Gly Arg Val
1 5 10 15
Val Arg Val Gly Glu Ile Val Arg Gly Glu Ala Lys Gly Val Trp Val
20 25 30
Val Ser Leu Asp Glu Ala Arg Leu Arg Leu Val Pro Arg Pro Val Val
35 40 45
Ala Ala Phe Pro Ser Gly Lys Ala Gln Val Tyr Ala Leu Arg Thr Ala
50 55 60
Thr Gly Arg Val Leu Glu Ala Thr Ala Asn His Pro Val Tyr Thr Pro
65 70 75 80
Glu Gly Trp Arg Pro Leu Gly Thr Leu Ala Pro Gly Asp Tyr Val Ala
85 90 95
Leu Pro Arg
<210> 32
<211> 40
<212> PRT
<213> Artificial Sequence
<400> 32
Met Ala Glu Val Tyr Trp Asp Arg Val Glu Ala Val Glu Pro Leu Gly
1 5 10 15
Glu Glu Glu Val Phe Asp Leu Thr Val Glu Gly Thr His Thr Phe Val
20 25 30
Ala Glu Asp Val Ile Val His Asn
35 40
<210> 33
<211> 14
<212> PRT
<213> Artificial Sequence
<400> 33
Met Ala His Ile Val Met Val Asp Ala Tyr Lys Pro Thr Lys
1 5 10
<210> 34
<211> 13
<212> PRT
<213> Artificial Sequence
<400> 34
Ala His Ile Val Met Val Asp Ala Tyr Lys Pro Thr Lys
1 5 10
<210> 35
<211> 117
<212> PRT
<213> Artificial Sequence
<400> 35
Met Gly Ala Met Val Asp Thr Leu Ser Gly Leu Ser Ser Glu Gln Gly
1 5 10 15
Gln Ser Gly Asp Met Thr Ile Glu Glu Asp Ser Ala Thr His Ile Lys
20 25 30
Phe Ser Lys Arg Asp Glu Asp Gly Lys Glu Leu Ala Gly Ala Thr Met
35 40 45
Glu Leu Arg Asp Ser Ser Gly Lys Thr Ile Ser Thr Trp Ile Ser Asp
50 55 60
Gly Gln Val Lys Asp Phe Tyr Leu Tyr Pro Gly Lys Tyr Thr Phe Val
65 70 75 80
Glu Thr Ala Ala Pro Asp Gly Tyr Glu Val Ala Thr Ala Ile Thr Phe
85 90 95
Thr Val Asn Glu Gln Gly Gln Val Thr Val Asn Gly Lys Ala Thr Lys
100 105 110
Gly Asp Ala His Ile
115
<210> 36
<211> 116
<212> PRT
<213> Artificial Sequence
<400> 36
Gly Ala Met Val Asp Thr Leu Ser Gly Leu Ser Ser Glu Gln Gly Gln
1 5 10 15
Ser Gly Asp Met Thr Ile Glu Glu Asp Ser Ala Thr His Ile Lys Phe
20 25 30
Ser Lys Arg Asp Glu Asp Gly Lys Glu Leu Ala Gly Ala Thr Met Glu
35 40 45
Leu Arg Asp Ser Ser Gly Lys Thr Ile Ser Thr Trp Ile Ser Asp Gly
50 55 60
Gln Val Lys Asp Phe Tyr Leu Tyr Pro Gly Lys Tyr Thr Phe Val Glu
65 70 75 80
Thr Ala Ala Pro Asp Gly Tyr Glu Val Ala Thr Ala Ile Thr Phe Thr
85 90 95
Val Asn Glu Gln Gly Gln Val Thr Val Asn Gly Lys Ala Thr Lys Gly
100 105 110
Asp Ala His Ile
115
<210> 37
<211> 34
<212> PRT
<213> Artificial Sequence
<400> 37
Met Arg Ala Lys Gln Leu Glu Asp Lys Ile Glu Glu Leu Leu Ser Lys
1 5 10 15
Ile Tyr His Leu Glu Asn Glu Ile Ala Arg Leu Lys Lys Leu Ile Gly
20 25 30
Glu Arg
<210> 38
<211> 28
<212> PRT
<213> Artificial Sequence
<400> 38
Gln Leu Glu Asp Lys Ile Glu Glu Leu Leu Ser Lys Ile Tyr His Leu
1 5 10 15
Glu Asn Glu Ile Ala Arg Leu Lys Lys Leu Ile Gly
20 25
<210> 39
<211> 34
<212> PRT
<213> Artificial Sequence
<400> 39
Met Arg Ala Lys Gln Leu Glu Asp Lys Val Glu Glu Leu Leu Ser Lys
1 5 10 15
Asn Tyr His Leu Glu Asn Glu Val Ala Arg Leu Lys Lys Leu Val Gly
20 25 30
Thr Arg
<210> 40
<211> 28
<212> PRT
<213> Artificial Sequence
<400> 40
Gln Leu Glu Asp Lys Val Glu Glu Leu Leu Ser Lys Asn Tyr His Leu
1 5 10 15
Glu Asn Glu Val Ala Arg Leu Lys Lys Leu Val Gly
20 25
<210> 41
<211> 16
<212> PRT
<213> Artificial Sequence
<400> 41
Ser Gly Gly Gly Ser Gly Gly Gly Ser Thr Pro Gln Ser Ala Phe Ala
1 5 10 15
<210> 42
<211> 11
<212> PRT
<213> Artificial Sequence
<400> 42
Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly
1 5 10
<210> 43
<211> 20
<212> PRT
<213> Artificial Sequence
<400> 43
Ser Gly Gly Ser Ser Gly Ser Thr Thr Thr Thr Arg Ile Thr Pro Gln
1 5 10 15
Ser Ala Phe Ala
20
<210> 44
<211> 12
<212> PRT
<213> Artificial Sequence
<400> 44
Gln Asn Thr Phe Ser Gln Gly Ser Ser Ser Gly Ser
1 5 10
<210> 45
<211> 4
<212> PRT
<213> Artificial Sequence
<400> 45
Ser Ala Phe Ala
1
<210> 46
<211> 36
<212> PRT
<213> Artificial Sequence
<400> 46
Gly Ala Ala Pro Ala Ala Ala Pro Ala Lys Gln Glu Ala Ala Ala Pro
1 5 10 15
Ala Pro Ala Ala Lys Ala Glu Ala Pro Ala Ala Ala Pro Ala Ala Lys
20 25 30
Ala Thr Pro Gln
35
<210> 47
<211> 33
<212> PRT
<213> Artificial Sequence
<400> 47
Ser Ala Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Glu Ala Ala
1 5 10 15
Ala Lys Glu Ala Ala Ala Lys Ala Val Asn Thr Pro Gln Ser Ala Phe
20 25 30
Ala
<210> 48
<211> 33
<212> PRT
<213> Artificial Sequence
<400> 48
Ser Ala Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Glu Ala Ala
1 5 10 15
Ala Lys Glu Ala Ala Ala Lys Ala Leu Asn Thr Pro Gln Ser Ala Phe
20 25 30
Ala
<210> 49
<211> 16
<212> PRT
<213> Artificial Sequence
<400> 49
Gly Gly Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Val Asn Leu
1 5 10 15
<210> 50
<211> 6
<212> PRT
<213> Artificial Sequence
<400> 50
Gly Ser Gly Gly Ser Gly
1 5
<210> 51
<211> 6
<212> PRT
<213> Artificial Sequence
<400> 51
Gly Ser Gly Gly Ser Gly
1 5
<210> 52
<211> 1233
<212> DNA
<213> Artificial Sequence
<400> 52
gcccaatcgg aaccggaact gaaactggaa agtgtggtta ttgtgtctcg tcatggcgtt 60
cgcgctccga ccaaatttac gcagctgatg caagatgtca ccccggacgc cttctatacg 120
tggccggtga agctgggtga actgaccccg cgtggcggtg aactgatcgc ctatctgggt 180
cactactggc gtcagcgcct ggtggcagat ggtctgctgc cgaaaaaggg ctgcccgcag 240
agcggtcaag ttgcaattat cgctgatgtc gacgaacgta cccgcaaaac gggtgaagca 300
tttgcggccg gtctggcacc ggattgcgcc attaccgttc atacgcaggc agataccagc 360
tctccggacc cgctgttcaa cccgctgaaa accggcgtct gtcagctgga tgtcgcgcaa 420
gtgacggacg ccattctgga acgtgcaggc ggttccatcg ctgattttac cggtcactac 480
cagacggcat tccgtgaact ggaacgcgtt ctgaactttc cgcagtcaaa tctggcgctg 540
aaacgcgaaa agcaggatga aagtgcgtcc ctgacccaag ccctgccgag tgaactgaaa 600
gtctccgccg acaatgtgtc actgaccggc gcatggtcac tggcttcgat gctgacggaa 660
atttttctgc tgcagcaagc acagggtatg ccggaaccgg gttggggtcg tatcaccgat 720
tcgcatcagt ggaacacgct gctgagcctg cacaatgcgc agttcgacct gctgcaacgt 780
accccggaag tggcacgttc gcgcgccacg ccgctgctgg atctgattaa aaccgctctg 840
acgccgcatc cgccgcagaa gcaagcgtat ggcgtgaccc tgccgacgag cgttctgttt 900
atcgcgggtc acgacaccaa cctggcaaat ctgggcggtg ctctggaact gcagtggacc 960
ctgccgggtc aaccggataa cacgccgccg ggcggtgaac tggttttcga acgttggcgt 1020
cgcctgagcg acaattctca gtggatccaa gttagcctgg tctttcagac cctgcagcaa 1080
atgcgcgata aaaccccgct gttcctgaac acgccgccgg gcgaagtgaa gctgaccctg 1140
gcgggttgcg aagaacgtaa cgcccagggc atgtgttctc tggcaggttt tacccagatt 1200
gttaatgaag cacgcatccc ggcttgtagt ctg 1233
<210> 53
<211> 411
<212> PRT
<213> Artificial Sequence
<400> 53
Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val Val Ile Val Ser
1 5 10 15
Arg His Gly Val Arg Ala Pro Thr Lys Phe Thr Gln Leu Met Gln Asp
20 25 30
Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro Val Lys Leu Gly Glu Leu
35 40 45
Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu Gly His Tyr Trp Arg
50 55 60
Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys Lys Gly Cys Pro Gln
65 70 75 80
Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp Glu Arg Thr Arg Lys
85 90 95
Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro Asp Cys Ala Ile Thr
100 105 110
Val His Thr Gln Ala Asp Thr Ser Ser Pro Asp Pro Leu Phe Asn Pro
115 120 125
Leu Lys Thr Gly Val Cys Gln Leu Asp Val Ala Gln Val Thr Asp Ala
130 135 140
Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp Phe Thr Gly His Tyr
145 150 155 160
Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu Asn Phe Pro Gln Ser
165 170 175
Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp Glu Ser Ala Ser Leu Thr
180 185 190
Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala Asp Asn Val Ser Leu
195 200 205
Thr Gly Ala Trp Ser Leu Ala Ser Met Leu Thr Glu Ile Phe Leu Leu
210 215 220
Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp Gly Arg Ile Thr Asp
225 230 235 240
Ser His Gln Trp Asn Thr Leu Leu Ser Leu His Asn Ala Gln Phe Asp
245 250 255
Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser Arg Ala Thr Pro Leu
260 265 270
Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His Pro Pro Gln Lys Gln
275 280 285
Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu Phe Ile Ala Gly His
290 295 300
Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu Glu Leu Gln Trp Thr
305 310 315 320
Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly Glu Leu Val Phe
325 330 335
Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln Trp Ile Gln Val Ser
340 345 350
Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp Lys Thr Pro Leu Phe
355 360 365
Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr Leu Ala Gly Cys Glu
370 375 380
Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala Gly Phe Thr Gln Ile
385 390 395 400
Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu
405 410
<210> 54
<211> 411
<212> PRT
<213> Artificial Sequence
<400> 54
Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val Val Ile Val Ser
1 5 10 15
Arg His Gly Val Arg Ala Pro Thr Lys Ala Thr Gln Leu Met Gln Asp
20 25 30
Val Thr Pro Asp Ala Trp Pro Thr Trp Pro Val Lys Leu Gly Glu Leu
35 40 45
Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu Gly His Tyr Trp Arg
50 55 60
Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys Cys Gly Cys Pro Gln
65 70 75 80
Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp Glu Arg Thr Arg Lys
85 90 95
Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro Asp Cys Ala Ile Thr
100 105 110
Val His Thr Gln Ala Asp Thr Ser Ser Pro Asp Pro Leu Phe Asn Pro
115 120 125
Leu Lys Thr Gly Val Cys Gln Leu Asp Asn Ala Asn Val Thr Asp Ala
130 135 140
Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp Phe Thr Gly His Tyr
145 150 155 160
Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu Asn Phe Pro Gln Ser
165 170 175
Asn Leu Cys Leu Lys Arg Glu Lys Gln Asp Glu Ser Cys Ser Leu Thr
180 185 190
Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala Asp Cys Val Ser Leu
195 200 205
Thr Gly Ala Val Ser Leu Ala Ser Met Leu Thr Glu Ile Phe Leu Leu
210 215 220
Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp Gly Arg Ile Thr Asp
225 230 235 240
Ser His Gln Trp Asn Thr Leu Leu Ser Leu His Asn Ala Gln Phe Asp
245 250 255
Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser Arg Ala Thr Pro Leu
260 265 270
Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His Pro Pro Gln Lys Gln
275 280 285
Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu Phe Ile Ala Gly His
290 295 300
Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu Glu Leu Asn Trp Thr
305 310 315 320
Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly Glu Leu Val Phe
325 330 335
Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln Trp Ile Gln Val Ser
340 345 350
Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp Lys Thr Pro Leu Ser
355 360 365
Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr Leu Ala Gly Cys Glu
370 375 380
Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala Gly Phe Thr Gln Ile
385 390 395 400
Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu
405 410
<210> 55
<211> 1254
<212> DNA
<213> Artificial Sequence
<400> 55
gcccagtcgg agcctgagct gaagctggag tcggtcgtca tcgtttcccg gcacggcgtt 60
agggccccaa ccaagttcac acagctgatg caggacgtga cccccgatgc ttggccaacc 120
tggcctgtca agctcggcga gctgaccccc aggggcgggg agctcatcgc gtacctgggg 180
cactactggc gccagcggct ggtcgctgac ggcctcctgc caaaggaagg ctgccctcag 240
tctgggcagg tggctatcat tgccgacgtc gatgagcgca ccaggaagac gggcgaggcc 300
ttcgccgcgg ggctcgcccc agactgcgct atcaccgtgc accatcaggc ggacacgtcc 360
agcccagatc ctctcttcaa cccgctgaag accggcgtct gccagctgga cgtggcgaat 420
gtccgcaggg ctatcctgga gagggccggc gggtccattg ccgatttcac aaggcattac 480
cagactgcgt tcagggagct ggagcgggtg ctgaacttcc cgcagagcaa tctctgcctg 540
aagcgcgaga agcaggacga gtcgtgctct ctcacgcagg ccctgccctc tgagctcaag 600
gtctcagcgg acgatgtttc cctgacaggc gcctggtcac tggcgtccat gctcactgag 660
attttcctcc tgcagcaggc tcaggggatg ccggagccag gctgggggcg gattacagac 720
agccaccagt ggaacactct cctgtcgctc cataatgcgg ttttcgatct cctgcagagg 780
acccccgagg tggctcggtc gcgcgccacg cccctcctgg acctgatcaa gacagctctc 840
actccacacc cgccccagaa gcaggcctac ggcgttaccc tgcctacgtc cgtgctcttc 900
attgccggcc atgataccaa cctcgctaat ctgggcgggg ccctggagct gaactggacc 960
ctgccgggcc agcccgacaa ttacccacct ggcggggagc tggtgttcga gaggtggagg 1020
cgcctcagcg ataactcgca gtggattcag gtgtccctcg tgttccagac actccagcag 1080
atgcgggaca agacaccgct ctcactgaac actccgcccg gcgaggtcaa gctcacgctg 1140
gccgggtgcg aggagaggaa cgctcagggc atgtgctccc tggctgggtt cacacagatt 1200
gtcaacgagg cccgcatccc cgcttgctct ctctccgaga aggacgagct gtaa 1254
<210> 56
<211> 411
<212> PRT
<213> Artificial Sequence
<400> 56
Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val Val Ile Val Ser
1 5 10 15
Arg His Gly Val Arg Ala Pro Thr Lys Phe Thr Gln Leu Met Gln Asp
20 25 30
Val Thr Pro Asp Ala Trp Pro Thr Trp Pro Val Lys Leu Gly Glu Leu
35 40 45
Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu Gly His Tyr Trp Arg
50 55 60
Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys Glu Gly Cys Pro Gln
65 70 75 80
Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp Glu Arg Thr Arg Lys
85 90 95
Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro Asp Cys Ala Ile Thr
100 105 110
Val His His Gln Ala Asp Thr Ser Ser Pro Asp Pro Leu Phe Asn Pro
115 120 125
Leu Lys Thr Gly Val Cys Gln Leu Asp Val Ala Asn Val Arg Arg Ala
130 135 140
Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp Phe Thr Arg His Tyr
145 150 155 160
Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu Asn Phe Pro Gln Ser
165 170 175
Asn Leu Cys Leu Lys Arg Glu Lys Gln Asp Glu Ser Cys Ser Leu Thr
180 185 190
Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala Asp Asp Val Ser Leu
195 200 205
Thr Gly Ala Trp Ser Leu Ala Ser Met Leu Thr Glu Ile Phe Leu Leu
210 215 220
Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp Gly Arg Ile Thr Asp
225 230 235 240
Ser His Gln Trp Asn Thr Leu Leu Ser Leu His Asn Ala Val Phe Asp
245 250 255
Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser Arg Ala Thr Pro Leu
260 265 270
Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His Pro Pro Gln Lys Gln
275 280 285
Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu Phe Ile Ala Gly His
290 295 300
Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu Glu Leu Asn Trp Thr
305 310 315 320
Leu Pro Gly Gln Pro Asp Asn Tyr Pro Pro Gly Gly Glu Leu Val Phe
325 330 335
Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln Trp Ile Gln Val Ser
340 345 350
Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp Lys Thr Pro Leu Ser
355 360 365
Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr Leu Ala Gly Cys Glu
370 375 380
Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala Gly Phe Thr Gln Ile
385 390 395 400
Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu
405 410
<210> 57
<211> 1641
<212> DNA
<213> Artificial Sequence
<400> 57
atgtcggacc tgttctggga taggatcgtg tcgattgagg agaaggggtc tgaggaggtc 60
tacgatctca cagttccaaa gtacgcttct tggctcgcgg atggggttgt ttcacataat 120
agcgcccaat cggaaccgga actgaaactg gaaagtgtgg ttattgtgtc tcgtcatggc 180
gttcgcgctc cgaccaaatt tacgcagctg atgcaagatg tcaccccgga cgccttctat 240
acgtggccgg tgaagctggg tgaactgacc ccgcgtggcg gtgaactgat cgcctatctg 300
ggtcactact ggcgtcagcg cctggtggca gatggtctgc tgccgaaaaa gggctgcccg 360
cagagcggtc aagttgcaat tatcgctgat gtcgacgaac gtacccgcaa aacgggtgaa 420
gcatttgcgg ccggtctggc accggattgc gccattaccg ttcatacgca ggcagatacc 480
agctctccgg acccgctgtt caacccgctg aaaaccggcg tctgtcagct ggatgtcgcg 540
caagtgacgg acgccattct ggaacgtgca ggcggttcca tcgctgattt taccggtcac 600
taccagacgg cattccgtga actggaacgc gttctgaact ttccgcagtc aaatctggcg 660
ctgaaacgcg aaaagcagga tgaaagtgcg tccctgaccc aagccctgcc gagtgaactg 720
aaagtctccg ccgacaatgt gtcactgacc ggcgcatggt cactggcttc gatgctgacg 780
gaaatttttc tgctgcagca agcacagggt atgccggaac cgggttgggg tcgtatcacc 840
gattcgcatc agtggaacac gctgctgagc ctgcacaatg cgcagttcga cctgctgcaa 900
cgtaccccgg aagtggcacg ttcgcgcgcc acgccgctgc tggatctgat taaaaccgct 960
ctgacgccgc atccgccgca gaagcaagcg tatggcgtga ccctgccgac gagcgttctg 1020
tttatcgcgg gtcacgacac caacctggca aatctgggcg gtgctctgga actgcagtgg 1080
accctgccgg gtcaaccgga taacacgccg ccgggcggtg aactggtttt cgaacgttgg 1140
cgtcgcctga gcgacaattc tcagtggatc caagttagcc tggtctttca gaccctgcag 1200
caaatgcgcg ataaaacccc gctgttcctg aacacgccgc cgggcgaagt gaagctgacc 1260
ctggcgggtt gcgaagaacg taacgcccag ggcatgtgtt ctctggcagg ttttacccag 1320
attgttaatg aagcacgcat cccggcttgt agtctgtgcg tgacagggga cactctcatc 1380
tgcctcgctg acgggcgccg cgttcctatt caggatctcg tggggcattc gccggaggtt 1440
attgcggtcg acgataaggg ccgcctcgtt tgcgctaagt cagaggtcat ctggaaggtc 1500
ggcgagcggt ccgttttcga gatcaagctg gcttccggga ggagcattaa ggctaccgct 1560
gagcacaggc tcctggcgtt caagggctgg aggcatgtta aggacttcaa agtgggggat 1620
aggctcgcta ttgctcacta a 1641
<210> 58
<211> 546
<212> PRT
<213> Artificial Sequence
<400> 58
Met Ser Asp Leu Phe Trp Asp Arg Ile Val Ser Ile Glu Glu Lys Gly
1 5 10 15
Ser Glu Glu Val Tyr Asp Leu Thr Val Pro Lys Tyr Ala Ser Trp Leu
20 25 30
Ala Asp Gly Val Val Ser His Asn Ser Ala Gln Ser Glu Pro Glu Leu
35 40 45
Lys Leu Glu Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala Pro
50 55 60
Thr Lys Phe Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr
65 70 75 80
Thr Trp Pro Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu
85 90 95
Ile Ala Tyr Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly
100 105 110
Leu Leu Pro Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile
115 120 125
Ala Asp Val Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala
130 135 140
Gly Leu Ala Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr
145 150 155 160
Ser Ser Pro Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln
165 170 175
Leu Asp Val Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly
180 185 190
Ser Ile Ala Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu
195 200 205
Glu Arg Val Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu
210 215 220
Lys Gln Asp Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu
225 230 235 240
Lys Val Ser Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala
245 250 255
Ser Met Leu Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro
260 265 270
Glu Pro Gly Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu
275 280 285
Leu Ser Leu His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu
290 295 300
Val Ala Arg Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala
305 310 315 320
Leu Thr Pro His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro
325 330 335
Thr Ser Val Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu
340 345 350
Gly Gly Ala Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn
355 360 365
Thr Pro Pro Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser
370 375 380
Asp Asn Ser Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln
385 390 395 400
Gln Met Arg Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu
405 410 415
Val Lys Leu Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met
420 425 430
Cys Ser Leu Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro
435 440 445
Ala Cys Ser Leu Cys Val Thr Gly Asp Thr Leu Ile Cys Leu Ala Asp
450 455 460
Gly Arg Arg Val Pro Ile Gln Asp Leu Val Gly His Ser Pro Glu Val
465 470 475 480
Ile Ala Val Asp Asp Lys Gly Arg Leu Val Cys Ala Lys Ser Glu Val
485 490 495
Ile Trp Lys Val Gly Glu Arg Ser Val Phe Glu Ile Lys Leu Ala Ser
500 505 510
Gly Arg Ser Ile Lys Ala Thr Ala Glu His Arg Leu Leu Ala Phe Lys
515 520 525
Gly Trp Arg His Val Lys Asp Phe Lys Val Gly Asp Arg Leu Ala Ile
530 535 540
Ala His
545
<210> 59
<211> 1650
<212> DNA
<213> Artificial Sequence
<400> 59
atggcggata tcgtttggac gaagttcaag attgaggagg tggagagcga tgttgagtat 60
gtgtacgatc tggaggtgga ggactaccac aacttcattg gcaatctcat catcaaccac 120
aacagcgccc aatcggaacc ggaactgaaa ctggaaagtg tggttattgt gtctcgtcat 180
ggcgttcgcg ctccgaccaa atttacgcag ctgatgcaag atgtcacccc ggacgccttc 240
tatacgtggc cggtgaagct gggtgaactg accccgcgtg gcggtgaact gatcgcctat 300
ctgggtcact actggcgtca gcgcctggtg gcagatggtc tgctgccgaa aaagggctgc 360
ccgcagagcg gtcaagttgc aattatcgct gatgtcgacg aacgtacccg caaaacgggt 420
gaagcatttg cggccggtct ggcaccggat tgcgccatta ccgttcatac gcaggcagat 480
accagctctc cggacccgct gttcaacccg ctgaaaaccg gcgtctgtca gctggatgtc 540
gcgcaagtga cggacgccat tctggaacgt gcaggcggtt ccatcgctga ttttaccggt 600
cactaccaga cggcattccg tgaactggaa cgcgttctga actttccgca gtcaaatctg 660
gcgctgaaac gcgaaaagca ggatgaaagt gcgtccctga cccaagccct gccgagtgaa 720
ctgaaagtct ccgccgacaa tgtgtcactg accggcgcat ggtcactggc ttcgatgctg 780
acggaaattt ttctgctgca gcaagcacag ggtatgccgg aaccgggttg gggtcgtatc 840
accgattcgc atcagtggaa cacgctgctg agcctgcaca atgcgcagtt cgacctgctg 900
caacgtaccc cggaagtggc acgttcgcgc gccacgccgc tgctggatct gattaaaacc 960
gctctgacgc cgcatccgcc gcagaagcaa gcgtatggcg tgaccctgcc gacgagcgtt 1020
ctgtttatcg cgggtcacga caccaacctg gcaaatctgg gcggtgctct ggaactgcag 1080
tggaccctgc cgggtcaacc ggataacacg ccgccgggcg gtgaactggt tttcgaacgt 1140
tggcgtcgcc tgagcgacaa ttctcagtgg atccaagtta gcctggtctt tcagaccctg 1200
cagcaaatgc gcgataaaac cccgctgttc ctgaacacgc cgccgggcga agtgaagctg 1260
accctggcgg gttgcgaaga acgtaacgcc cagggcatgt gttctctggc aggttttacc 1320
cagattgtta atgaagcacg catcccggct tgtagtctgt gcctgcaccc tgacacatac 1380
gttattctcc ctgacgggcg catgaagaag atttcggaga ttgatgagga tgaggttctc 1440
tcagtcaact tcgaggacct gaagctctac aataagaaga tcaagaagtt caagcacaag 1500
gctccgaaga tcctctacaa gattaagacc gcgttctccg agctcatcac cacgggcgag 1560
cataagctgt tcgtggtcga gaacgggaag atcgtcgaga agtgcgttaa ggacctcaat 1620
ggcagcgagc tgatcggggt tgtgaggtaa 1650
<210> 60
<211> 549
<212> PRT
<213> Artificial Sequence
<400> 60
Met Ala Asp Ile Val Trp Thr Lys Phe Lys Ile Glu Glu Val Glu Ser
1 5 10 15
Asp Val Glu Tyr Val Tyr Asp Leu Glu Val Glu Asp Tyr His Asn Phe
20 25 30
Ile Gly Asn Leu Ile Ile Asn His Asn Ser Ala Gln Ser Glu Pro Glu
35 40 45
Leu Lys Leu Glu Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala
50 55 60
Pro Thr Lys Phe Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe
65 70 75 80
Tyr Thr Trp Pro Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu
85 90 95
Leu Ile Ala Tyr Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp
100 105 110
Gly Leu Leu Pro Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile
115 120 125
Ile Ala Asp Val Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala
130 135 140
Ala Gly Leu Ala Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp
145 150 155 160
Thr Ser Ser Pro Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys
165 170 175
Gln Leu Asp Val Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly
180 185 190
Gly Ser Ile Ala Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu
195 200 205
Leu Glu Arg Val Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg
210 215 220
Glu Lys Gln Asp Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu
225 230 235 240
Leu Lys Val Ser Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu
245 250 255
Ala Ser Met Leu Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met
260 265 270
Pro Glu Pro Gly Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr
275 280 285
Leu Leu Ser Leu His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro
290 295 300
Glu Val Ala Arg Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr
305 310 315 320
Ala Leu Thr Pro His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu
325 330 335
Pro Thr Ser Val Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn
340 345 350
Leu Gly Gly Ala Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp
355 360 365
Asn Thr Pro Pro Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu
370 375 380
Ser Asp Asn Ser Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu
385 390 395 400
Gln Gln Met Arg Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly
405 410 415
Glu Val Lys Leu Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly
420 425 430
Met Cys Ser Leu Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile
435 440 445
Pro Ala Cys Ser Leu Cys Leu His Pro Asp Thr Tyr Val Ile Leu Pro
450 455 460
Asp Gly Arg Met Lys Lys Ile Ser Glu Ile Asp Glu Asp Glu Val Leu
465 470 475 480
Ser Val Asn Phe Glu Asp Leu Lys Leu Tyr Asn Lys Lys Ile Lys Lys
485 490 495
Phe Lys His Lys Ala Pro Lys Ile Leu Tyr Lys Ile Lys Thr Ala Phe
500 505 510
Ser Glu Leu Ile Thr Thr Gly Glu His Lys Leu Phe Val Val Glu Asn
515 520 525
Gly Lys Ile Val Glu Lys Cys Val Lys Asp Leu Asn Gly Ser Glu Leu
530 535 540
Ile Gly Val Val Arg
545
<210> 61
<211> 1740
<212> DNA
<213> Artificial Sequence
<400> 61
atggggaatt acctgtacgc tcccatcatt aggatcggcc gggagtacta cgacgggttc 60
gtctacaatc tggaggtgga ggatgactct tcatacgtta cagtctcagg cactctgcac 120
aacagcgccc aatcggaacc ggaactgaaa ctggaaagtg tggttattgt gtctcgtcat 180
ggcgttcgcg ctccgaccaa atttacgcag ctgatgcaag atgtcacccc ggacgccttc 240
tatacgtggc cggtgaagct gggtgaactg accccgcgtg gcggtgaact gatcgcctat 300
ctgggtcact actggcgtca gcgcctggtg gcagatggtc tgctgccgaa aaagggctgc 360
ccgcagagcg gtcaagttgc aattatcgct gatgtcgacg aacgtacccg caaaacgggt 420
gaagcatttg cggccggtct ggcaccggat tgcgccatta ccgttcatac gcaggcagat 480
accagctctc cggacccgct gttcaacccg ctgaaaaccg gcgtctgtca gctggatgtc 540
gcgcaagtga cggacgccat tctggaacgt gcaggcggtt ccatcgctga ttttaccggt 600
cactaccaga cggcattccg tgaactggaa cgcgttctga actttccgca gtcaaatctg 660
gcgctgaaac gcgaaaagca ggatgaaagt gcgtccctga cccaagccct gccgagtgaa 720
ctgaaagtct ccgccgacaa tgtgtcactg accggcgcat ggtcactggc ttcgatgctg 780
acggaaattt ttctgctgca gcaagcacag ggtatgccgg aaccgggttg gggtcgtatc 840
accgattcgc atcagtggaa cacgctgctg agcctgcaca atgcgcagtt cgacctgctg 900
caacgtaccc cggaagtggc acgttcgcgc gccacgccgc tgctggatct gattaaaacc 960
gctctgacgc cgcatccgcc gcagaagcaa gcgtatggcg tgaccctgcc gacgagcgtt 1020
ctgtttatcg cgggtcacga caccaacctg gcaaatctgg gcggtgctct ggaactgcag 1080
tggaccctgc cgggtcaacc ggataacacg ccgccgggcg gtgaactggt tttcgaacgt 1140
tggcgtcgcc tgagcgacaa ttctcagtgg atccaagtta gcctggtctt tcagaccctg 1200
cagcaaatgc gcgataaaac cccgctgttc ctgaacacgc cgccgggcga agtgaagctg 1260
accctggcgg gttgcgaaga acgtaacgcc cagggcatgt gttctctggc aggttttacc 1320
cagattgtta atgaagcacg catcccggct tgtagtctgt gcgttccgcc tgacactctg 1380
ctcatcctgg agaatgggtt caagcgcatc gtggacatta aggtcgggga caaggtcctg 1440
acgcacgaga accggttcaa gaaggttgag aaggtgtaca agcgcaggta catcggcgac 1500
atcattaaga ttaaggtgcg ctacttccca gaggagatca ttctcacccc agagcaccct 1560
gtctacgcta tcaagacgga gaagaggtgc gatggctctc atgggatctg caagttcaac 1620
tgcctcacac agtacactaa tccttcatgc aagaagcggt accgcaagta caagagggag 1680
tggatcattg ccaaggacct gaaggtcggc gatgtgatcg tctacccgat tcccaactaa 1740
<210> 62
<211> 579
<212> PRT
<213> Artificial Sequence
<400> 62
Met Gly Asn Tyr Leu Tyr Ala Pro Ile Ile Arg Ile Gly Arg Glu Tyr
1 5 10 15
Tyr Asp Gly Phe Val Tyr Asn Leu Glu Val Glu Asp Asp Ser Ser Tyr
20 25 30
Val Thr Val Ser Gly Thr Leu His Asn Ser Ala Gln Ser Glu Pro Glu
35 40 45
Leu Lys Leu Glu Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala
50 55 60
Pro Thr Lys Phe Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe
65 70 75 80
Tyr Thr Trp Pro Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu
85 90 95
Leu Ile Ala Tyr Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp
100 105 110
Gly Leu Leu Pro Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile
115 120 125
Ile Ala Asp Val Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala
130 135 140
Ala Gly Leu Ala Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp
145 150 155 160
Thr Ser Ser Pro Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys
165 170 175
Gln Leu Asp Val Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly
180 185 190
Gly Ser Ile Ala Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu
195 200 205
Leu Glu Arg Val Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg
210 215 220
Glu Lys Gln Asp Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu
225 230 235 240
Leu Lys Val Ser Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu
245 250 255
Ala Ser Met Leu Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met
260 265 270
Pro Glu Pro Gly Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr
275 280 285
Leu Leu Ser Leu His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro
290 295 300
Glu Val Ala Arg Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr
305 310 315 320
Ala Leu Thr Pro His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu
325 330 335
Pro Thr Ser Val Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn
340 345 350
Leu Gly Gly Ala Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp
355 360 365
Asn Thr Pro Pro Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu
370 375 380
Ser Asp Asn Ser Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu
385 390 395 400
Gln Gln Met Arg Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly
405 410 415
Glu Val Lys Leu Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly
420 425 430
Met Cys Ser Leu Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile
435 440 445
Pro Ala Cys Ser Leu Cys Val Pro Pro Asp Thr Leu Leu Ile Leu Glu
450 455 460
Asn Gly Phe Lys Arg Ile Val Asp Ile Lys Val Gly Asp Lys Val Leu
465 470 475 480
Thr His Glu Asn Arg Phe Lys Lys Val Glu Lys Val Tyr Lys Arg Arg
485 490 495
Tyr Ile Gly Asp Ile Ile Lys Ile Lys Val Arg Tyr Phe Pro Glu Glu
500 505 510
Ile Ile Leu Thr Pro Glu His Pro Val Tyr Ala Ile Lys Thr Glu Lys
515 520 525
Arg Cys Asp Gly Ser His Gly Ile Cys Lys Phe Asn Cys Leu Thr Gln
530 535 540
Tyr Thr Asn Pro Ser Cys Lys Lys Arg Tyr Arg Lys Tyr Lys Arg Glu
545 550 555 560
Trp Ile Ile Ala Lys Asp Leu Lys Val Gly Asp Val Ile Val Tyr Pro
565 570 575
Ile Pro Asn
<210> 63
<211> 1686
<212> DNA
<213> Artificial Sequence
<400> 63
atgaacattg cgttcgtcga ggttgaggat gtcgagatca ttgactacga tggctacgtt 60
tacgatctca caacagagac tcataacttc attgctaatg gcatcgtggt tcataatagc 120
gcccaatcgg aaccggaact gaaactggaa agtgtggtta ttgtgtctcg tcatggcgtt 180
cgcgctccga ccaaatttac gcagctgatg caagatgtca ccccggacgc cttctatacg 240
tggccggtga agctgggtga actgaccccg cgtggcggtg aactgatcgc ctatctgggt 300
cactactggc gtcagcgcct ggtggcagat ggtctgctgc cgaaaaaggg ctgcccgcag 360
agcggtcaag ttgcaattat cgctgatgtc gacgaacgta cccgcaaaac gggtgaagca 420
tttgcggccg gtctggcacc ggattgcgcc attaccgttc atacgcaggc agataccagc 480
tctccggacc cgctgttcaa cccgctgaaa accggcgtct gtcagctgga tgtcgcgcaa 540
gtgacggacg ccattctgga acgtgcaggc ggttccatcg ctgattttac cggtcactac 600
cagacggcat tccgtgaact ggaacgcgtt ctgaactttc cgcagtcaaa tctggcgctg 660
aaacgcgaaa agcaggatga aagtgcgtcc ctgacccaag ccctgccgag tgaactgaaa 720
gtctccgccg acaatgtgtc actgaccggc gcatggtcac tggcttcgat gctgacggaa 780
atttttctgc tgcagcaagc acagggtatg ccggaaccgg gttggggtcg tatcaccgat 840
tcgcatcagt ggaacacgct gctgagcctg cacaatgcgc agttcgacct gctgcaacgt 900
accccggaag tggcacgttc gcgcgccacg ccgctgctgg atctgattaa aaccgctctg 960
acgccgcatc cgccgcagaa gcaagcgtat ggcgtgaccc tgccgacgag cgttctgttt 1020
atcgcgggtc acgacaccaa cctggcaaat ctgggcggtg ctctggaact gcagtggacc 1080
ctgccgggtc aaccggataa cacgccgccg ggcggtgaac tggttttcga acgttggcgt 1140
cgcctgagcg acaattctca gtggatccaa gttagcctgg tctttcagac cctgcagcaa 1200
atgcgcgata aaaccccgct gttcctgaac acgccgccgg gcgaagtgaa gctgaccctg 1260
gcgggttgcg aagaacgtaa cgcccagggc atgtgttctc tggcaggttt tacccagatt 1320
gttaatgaag cacgcatccc ggcttgtagt ctgtgcctga tgccgcatga gaaggtgctg 1380
acggagtacg gggagattaa gattgaggac ctgttcaaga tcgggaagga gatcgtggag 1440
aaggacgagc tcaaggagat caggaagctg aatattaagg tgcacactct caacgagaat 1500
ggcgagatca agatcattaa cgccccatac gtgtggaagc tcaagcataa ggggaagatg 1560
atcaaggtca agctgaagaa ctggcactcg atcaccacga caccggagca tcccttcctg 1620
accaacaatg gctggatcaa ggcggagaat attaagaagg ggatgtatgt ggctatccct 1680
cgctaa 1686
<210> 64
<211> 561
<212> PRT
<213> Artificial Sequence
<400> 64
Met Asn Ile Ala Phe Val Glu Val Glu Asp Val Glu Ile Ile Asp Tyr
1 5 10 15
Asp Gly Tyr Val Tyr Asp Leu Thr Thr Glu Thr His Asn Phe Ile Ala
20 25 30
Asn Gly Ile Val Val His Asn Ser Ala Gln Ser Glu Pro Glu Leu Lys
35 40 45
Leu Glu Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr
50 55 60
Lys Phe Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr
65 70 75 80
Trp Pro Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile
85 90 95
Ala Tyr Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu
100 105 110
Leu Pro Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala
115 120 125
Asp Val Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly
130 135 140
Leu Ala Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser
145 150 155 160
Ser Pro Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu
165 170 175
Asp Val Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser
180 185 190
Ile Ala Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu
195 200 205
Arg Val Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys
210 215 220
Gln Asp Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys
225 230 235 240
Val Ser Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser
245 250 255
Met Leu Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu
260 265 270
Pro Gly Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu
275 280 285
Ser Leu His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val
290 295 300
Ala Arg Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu
305 310 315 320
Thr Pro His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr
325 330 335
Ser Val Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly
340 345 350
Gly Ala Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr
355 360 365
Pro Pro Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp
370 375 380
Asn Ser Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln
385 390 395 400
Met Arg Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val
405 410 415
Lys Leu Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys
420 425 430
Ser Leu Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala
435 440 445
Cys Ser Leu Cys Leu Met Pro His Glu Lys Val Leu Thr Glu Tyr Gly
450 455 460
Glu Ile Lys Ile Glu Asp Leu Phe Lys Ile Gly Lys Glu Ile Val Glu
465 470 475 480
Lys Asp Glu Leu Lys Glu Ile Arg Lys Leu Asn Ile Lys Val His Thr
485 490 495
Leu Asn Glu Asn Gly Glu Ile Lys Ile Ile Asn Ala Pro Tyr Val Trp
500 505 510
Lys Leu Lys His Lys Gly Lys Met Ile Lys Val Lys Leu Lys Asn Trp
515 520 525
His Ser Ile Thr Thr Thr Pro Glu His Pro Phe Leu Thr Asn Asn Gly
530 535 540
Trp Ile Lys Ala Glu Asn Ile Lys Lys Gly Met Tyr Val Ala Ile Pro
545 550 555 560
Arg
<210> 65
<211> 1668
<212> DNA
<213> Artificial Sequence
<400> 65
atgtacgggt tctacgacct cgacgatgtg tgcgtctcac tggagtccta caagggcgag 60
gtgtacgatc tcactctgga gggcaggcct tactacttcg ccaatggcat cctcactcat 120
aatagcgccc aatcggaacc ggaactgaaa ctggaaagtg tggttattgt gtctcgtcat 180
ggcgttcgcg ctccgaccaa atttacgcag ctgatgcaag atgtcacccc ggacgccttc 240
tatacgtggc cggtgaagct gggtgaactg accccgcgtg gcggtgaact gatcgcctat 300
ctgggtcact actggcgtca gcgcctggtg gcagatggtc tgctgccgaa aaagggctgc 360
ccgcagagcg gtcaagttgc aattatcgct gatgtcgacg aacgtacccg caaaacgggt 420
gaagcatttg cggccggtct ggcaccggat tgcgccatta ccgttcatac gcaggcagat 480
accagctctc cggacccgct gttcaacccg ctgaaaaccg gcgtctgtca gctggatgtc 540
gcgcaagtga cggacgccat tctggaacgt gcaggcggtt ccatcgctga ttttaccggt 600
cactaccaga cggcattccg tgaactggaa cgcgttctga actttccgca gtcaaatctg 660
gcgctgaaac gcgaaaagca ggatgaaagt gcgtccctga cccaagccct gccgagtgaa 720
ctgaaagtct ccgccgacaa tgtgtcactg accggcgcat ggtcactggc ttcgatgctg 780
acggaaattt ttctgctgca gcaagcacag ggtatgccgg aaccgggttg gggtcgtatc 840
accgattcgc atcagtggaa cacgctgctg agcctgcaca atgcgcagtt cgacctgctg 900
caacgtaccc cggaagtggc acgttcgcgc gccacgccgc tgctggatct gattaaaacc 960
gctctgacgc cgcatccgcc gcagaagcaa gcgtatggcg tgaccctgcc gacgagcgtt 1020
ctgtttatcg cgggtcacga caccaacctg gcaaatctgg gcggtgctct ggaactgcag 1080
tggaccctgc cgggtcaacc ggataacacg ccgccgggcg gtgaactggt tttcgaacgt 1140
tggcgtcgcc tgagcgacaa ttctcagtgg atccaagtta gcctggtctt tcagaccctg 1200
cagcaaatgc gcgataaaac cccgctgttc ctgaacacgc cgccgggcga agtgaagctg 1260
accctggcgg gttgcgaaga acgtaacgcc cagggcatgt gttctctggc aggttttacc 1320
cagattgtta atgaagcacg catcccggct tgtagtctgt gccatccaaa ggggacaaag 1380
gtcgtggtca agggcaaggg catcgtgaat attgaggacg ttaaggaggg gaattacgtt 1440
ctcggcatcg acggctggca gaaggttaag aaggtctgga agtacgagta cgagggcgag 1500
ctcattaacg ttaatgggct gaagtgcaca ccgaaccaca agatccccct ccgctacaag 1560
attaagcata agaagatcaa caagaacgat tacctggtga gggacatcta cgcgaagtcg 1620
ctcctgacca agttcaaggg cgaggggaag ctcatcctgt gcaagtaa 1668
<210> 66
<211> 555
<212> PRT
<213> Artificial Sequence
<400> 66
Met Tyr Gly Phe Tyr Asp Leu Asp Asp Val Cys Val Ser Leu Glu Ser
1 5 10 15
Tyr Lys Gly Glu Val Tyr Asp Leu Thr Leu Glu Gly Arg Pro Tyr Tyr
20 25 30
Phe Ala Asn Gly Ile Leu Thr His Asn Ser Ala Gln Ser Glu Pro Glu
35 40 45
Leu Lys Leu Glu Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala
50 55 60
Pro Thr Lys Phe Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe
65 70 75 80
Tyr Thr Trp Pro Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu
85 90 95
Leu Ile Ala Tyr Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp
100 105 110
Gly Leu Leu Pro Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile
115 120 125
Ile Ala Asp Val Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala
130 135 140
Ala Gly Leu Ala Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp
145 150 155 160
Thr Ser Ser Pro Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys
165 170 175
Gln Leu Asp Val Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly
180 185 190
Gly Ser Ile Ala Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu
195 200 205
Leu Glu Arg Val Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg
210 215 220
Glu Lys Gln Asp Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu
225 230 235 240
Leu Lys Val Ser Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu
245 250 255
Ala Ser Met Leu Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met
260 265 270
Pro Glu Pro Gly Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr
275 280 285
Leu Leu Ser Leu His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro
290 295 300
Glu Val Ala Arg Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr
305 310 315 320
Ala Leu Thr Pro His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu
325 330 335
Pro Thr Ser Val Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn
340 345 350
Leu Gly Gly Ala Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp
355 360 365
Asn Thr Pro Pro Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu
370 375 380
Ser Asp Asn Ser Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu
385 390 395 400
Gln Gln Met Arg Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly
405 410 415
Glu Val Lys Leu Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly
420 425 430
Met Cys Ser Leu Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile
435 440 445
Pro Ala Cys Ser Leu Cys His Pro Lys Gly Thr Lys Val Val Val Lys
450 455 460
Gly Lys Gly Ile Val Asn Ile Glu Asp Val Lys Glu Gly Asn Tyr Val
465 470 475 480
Leu Gly Ile Asp Gly Trp Gln Lys Val Lys Lys Val Trp Lys Tyr Glu
485 490 495
Tyr Glu Gly Glu Leu Ile Asn Val Asn Gly Leu Lys Cys Thr Pro Asn
500 505 510
His Lys Ile Pro Leu Arg Tyr Lys Ile Lys His Lys Lys Ile Asn Lys
515 520 525
Asn Asp Tyr Leu Val Arg Asp Ile Tyr Ala Lys Ser Leu Leu Thr Lys
530 535 540
Phe Lys Gly Glu Gly Lys Leu Ile Leu Cys Lys
545 550 555
<210> 67
<211> 1728
<212> DNA
<213> Artificial Sequence
<400> 67
atgtccgtga gctgggacga ggtcgcggag atcctggagt acgagccaaa ggatccttgg 60
gtctacgatc tgcaggttcc aggctaccac aacttcctcg ctaatggcat cttcgttcat 120
aatagcgccc aatcggaacc ggaactgaaa ctggaaagtg tggttattgt gtctcgtcat 180
ggcgttcgcg ctccgaccaa atttacgcag ctgatgcaag atgtcacccc ggacgccttc 240
tatacgtggc cggtgaagct gggtgaactg accccgcgtg gcggtgaact gatcgcctat 300
ctgggtcact actggcgtca gcgcctggtg gcagatggtc tgctgccgaa aaagggctgc 360
ccgcagagcg gtcaagttgc aattatcgct gatgtcgacg aacgtacccg caaaacgggt 420
gaagcatttg cggccggtct ggcaccggat tgcgccatta ccgttcatac gcaggcagat 480
accagctctc cggacccgct gttcaacccg ctgaaaaccg gcgtctgtca gctggatgtc 540
gcgcaagtga cggacgccat tctggaacgt gcaggcggtt ccatcgctga ttttaccggt 600
cactaccaga cggcattccg tgaactggaa cgcgttctga actttccgca gtcaaatctg 660
gcgctgaaac gcgaaaagca ggatgaaagt gcgtccctga cccaagccct gccgagtgaa 720
ctgaaagtct ccgccgacaa tgtgtcactg accggcgcat ggtcactggc ttcgatgctg 780
acggaaattt ttctgctgca gcaagcacag ggtatgccgg aaccgggttg gggtcgtatc 840
accgattcgc atcagtggaa cacgctgctg agcctgcaca atgcgcagtt cgacctgctg 900
caacgtaccc cggaagtggc acgttcgcgc gccacgccgc tgctggatct gattaaaacc 960
gctctgacgc cgcatccgcc gcagaagcaa gcgtatggcg tgaccctgcc gacgagcgtt 1020
ctgtttatcg cgggtcacga caccaacctg gcaaatctgg gcggtgctct ggaactgcag 1080
tggaccctgc cgggtcaacc ggataacacg ccgccgggcg gtgaactggt tttcgaacgt 1140
tggcgtcgcc tgagcgacaa ttctcagtgg atccaagtta gcctggtctt tcagaccctg 1200
cagcaaatgc gcgataaaac cccgctgttc ctgaacacgc cgccgggcga agtgaagctg 1260
accctggcgg gttgcgaaga acgtaacgcc cagggcatgt gttctctggc aggttttacc 1320
cagattgtta atgaagcacg catcccggct tgtagtctgt gcgtggatta cgagactgag 1380
gtcgtgctgg ggaatgggga gcggaagaag atcggggaga tcgtggagcg ggctattgag 1440
gaggctgaga agaacggcaa gctcgggcgg gttgacgatg gcttctacgc tccgatcgac 1500
attgaggtct actcgctcga tctggagacc ctcaaggttc ggaaggcgcg ggcaaatatc 1560
gcgtggaagc gcacagctcc aaagaagatg atgctggtga agactagggg cgggaagcgc 1620
attagggtca ccccgacgca ccccttcttc gttctggagg agggcaaggt ggctatgagg 1680
aaggcccggg acctggagga gggcaacaag atcgccacga ttgagtaa 1728
<210> 68
<211> 575
<212> PRT
<213> Artificial Sequence
<400> 68
Met Ser Val Ser Trp Asp Glu Val Ala Glu Ile Leu Glu Tyr Glu Pro
1 5 10 15
Lys Asp Pro Trp Val Tyr Asp Leu Gln Val Pro Gly Tyr His Asn Phe
20 25 30
Leu Ala Asn Gly Ile Phe Val His Asn Ser Ala Gln Ser Glu Pro Glu
35 40 45
Leu Lys Leu Glu Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala
50 55 60
Pro Thr Lys Phe Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe
65 70 75 80
Tyr Thr Trp Pro Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu
85 90 95
Leu Ile Ala Tyr Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp
100 105 110
Gly Leu Leu Pro Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile
115 120 125
Ile Ala Asp Val Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala
130 135 140
Ala Gly Leu Ala Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp
145 150 155 160
Thr Ser Ser Pro Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys
165 170 175
Gln Leu Asp Val Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly
180 185 190
Gly Ser Ile Ala Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu
195 200 205
Leu Glu Arg Val Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg
210 215 220
Glu Lys Gln Asp Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu
225 230 235 240
Leu Lys Val Ser Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu
245 250 255
Ala Ser Met Leu Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met
260 265 270
Pro Glu Pro Gly Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr
275 280 285
Leu Leu Ser Leu His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro
290 295 300
Glu Val Ala Arg Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr
305 310 315 320
Ala Leu Thr Pro His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu
325 330 335
Pro Thr Ser Val Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn
340 345 350
Leu Gly Gly Ala Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp
355 360 365
Asn Thr Pro Pro Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu
370 375 380
Ser Asp Asn Ser Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu
385 390 395 400
Gln Gln Met Arg Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly
405 410 415
Glu Val Lys Leu Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly
420 425 430
Met Cys Ser Leu Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile
435 440 445
Pro Ala Cys Ser Leu Cys Val Asp Tyr Glu Thr Glu Val Val Leu Gly
450 455 460
Asn Gly Glu Arg Lys Lys Ile Gly Glu Ile Val Glu Arg Ala Ile Glu
465 470 475 480
Glu Ala Glu Lys Asn Gly Lys Leu Gly Arg Val Asp Asp Gly Phe Tyr
485 490 495
Ala Pro Ile Asp Ile Glu Val Tyr Ser Leu Asp Leu Glu Thr Leu Lys
500 505 510
Val Arg Lys Ala Arg Ala Asn Ile Ala Trp Lys Arg Thr Ala Pro Lys
515 520 525
Lys Met Met Leu Val Lys Thr Arg Gly Gly Lys Arg Ile Arg Val Thr
530 535 540
Pro Thr His Pro Phe Phe Val Leu Glu Glu Gly Lys Val Ala Met Arg
545 550 555 560
Lys Ala Arg Asp Leu Glu Glu Gly Asn Lys Ile Ala Thr Ile Glu
565 570 575
<210> 69
<211> 1689
<212> DNA
<213> Artificial Sequence
<400> 69
atgacgctgg tgttcatccc cgttgagaat gtggaggagg aggagtacga cggctacgtt 60
tacgatctca ctacggagac tcataacttc attgctaatg gcatcctcgt tcataatagc 120
gcccaatcgg aaccggaact gaaactggaa agtgtggtta ttgtgtctcg tcatggcgtt 180
cgcgctccga ccaaatttac gcagctgatg caagatgtca ccccggacgc cttctatacg 240
tggccggtga agctgggtga actgaccccg cgtggcggtg aactgatcgc ctatctgggt 300
cactactggc gtcagcgcct ggtggcagat ggtctgctgc cgaaaaaggg ctgcccgcag 360
agcggtcaag ttgcaattat cgctgatgtc gacgaacgta cccgcaaaac gggtgaagca 420
tttgcggccg gtctggcacc ggattgcgcc attaccgttc atacgcaggc agataccagc 480
tctccggacc cgctgttcaa cccgctgaaa accggcgtct gtcagctgga tgtcgcgcaa 540
gtgacggacg ccattctgga acgtgcaggc ggttccatcg ctgattttac cggtcactac 600
cagacggcat tccgtgaact ggaacgcgtt ctgaactttc cgcagtcaaa tctggcgctg 660
aaacgcgaaa agcaggatga aagtgcgtcc ctgacccaag ccctgccgag tgaactgaaa 720
gtctccgccg acaatgtgtc actgaccggc gcatggtcac tggcttcgat gctgacggaa 780
atttttctgc tgcagcaagc acagggtatg ccggaaccgg gttggggtcg tatcaccgat 840
tcgcatcagt ggaacacgct gctgagcctg cacaatgcgc agttcgacct gctgcaacgt 900
accccggaag tggcacgttc gcgcgccacg ccgctgctgg atctgattaa aaccgctctg 960
acgccgcatc cgccgcagaa gcaagcgtat ggcgtgaccc tgccgacgag cgttctgttt 1020
atcgcgggtc acgacaccaa cctggcaaat ctgggcggtg ctctggaact gcagtggacc 1080
ctgccgggtc aaccggataa cacgccgccg ggcggtgaac tggttttcga acgttggcgt 1140
cgcctgagcg acaattctca gtggatccaa gttagcctgg tctttcagac cctgcagcaa 1200
atgcgcgata aaaccccgct gttcctgaac acgccgccgg gcgaagtgaa gctgaccctg 1260
gcgggttgcg aagaacgtaa cgcccagggc atgtgttctc tggcaggttt tacccagatt 1320
gttaatgaag cacgcatccc ggcttgtagt ctgtgcctcc tccctgatga gaaggtcgtg 1380
gttccctcgg tcgggttcgt gacactcaag gagctgttcg agacggcttc caaggtcgtg 1440
gagcgcgacg atgagaagga gatcagggag ctcgacgagc ggattaccag cgttaacggc 1500
gatgggaaga cgggcctggt caaggcctcc tacgtgtgga aggttaggca caagggcaag 1560
gtcatccggg tcaagctcaa gaattggcac ggcgttacag tgactccgga gcatcccttc 1620
ctcaccacga aggggtggaa gagggctgac cagctgaggc caggcgatta cgtcgcggtt 1680
cctaggtaa 1689
<210> 70
<211> 562
<212> PRT
<213> Artificial Sequence
<400> 70
Met Thr Leu Val Phe Ile Pro Val Glu Asn Val Glu Glu Glu Glu Tyr
1 5 10 15
Asp Gly Tyr Val Tyr Asp Leu Thr Thr Glu Thr His Asn Phe Ile Ala
20 25 30
Asn Gly Ile Leu Val His Asn Ser Ala Gln Ser Glu Pro Glu Leu Lys
35 40 45
Leu Glu Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr
50 55 60
Lys Phe Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr
65 70 75 80
Trp Pro Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile
85 90 95
Ala Tyr Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu
100 105 110
Leu Pro Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala
115 120 125
Asp Val Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly
130 135 140
Leu Ala Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser
145 150 155 160
Ser Pro Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu
165 170 175
Asp Val Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser
180 185 190
Ile Ala Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu
195 200 205
Arg Val Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys
210 215 220
Gln Asp Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys
225 230 235 240
Val Ser Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser
245 250 255
Met Leu Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu
260 265 270
Pro Gly Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu
275 280 285
Ser Leu His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val
290 295 300
Ala Arg Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu
305 310 315 320
Thr Pro His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr
325 330 335
Ser Val Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly
340 345 350
Gly Ala Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr
355 360 365
Pro Pro Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp
370 375 380
Asn Ser Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln
385 390 395 400
Met Arg Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val
405 410 415
Lys Leu Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys
420 425 430
Ser Leu Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala
435 440 445
Cys Ser Leu Cys Leu Leu Pro Asp Glu Lys Val Val Val Pro Ser Val
450 455 460
Gly Phe Val Thr Leu Lys Glu Leu Phe Glu Thr Ala Ser Lys Val Val
465 470 475 480
Glu Arg Asp Asp Glu Lys Glu Ile Arg Glu Leu Asp Glu Arg Ile Thr
485 490 495
Ser Val Asn Gly Asp Gly Lys Thr Gly Leu Val Lys Ala Ser Tyr Val
500 505 510
Trp Lys Val Arg His Lys Gly Lys Val Ile Arg Val Lys Leu Lys Asn
515 520 525
Trp His Gly Val Thr Val Thr Pro Glu His Pro Phe Leu Thr Thr Lys
530 535 540
Gly Trp Lys Arg Ala Asp Gln Leu Arg Pro Gly Asp Tyr Val Ala Val
545 550 555 560
Pro Arg
<210> 71
<211> 1728
<212> DNA
<213> Artificial Sequence
<400> 71
atgacccatg ttctgttcga cgagatcgtg gagattcggt acatctccga gggccaggag 60
gtgtacgacg ttactacgga gactcataat ttcattgggg gcaacatgcc tactctgctc 120
cacaacagcg cccaatcgga accggaactg aaactggaaa gtgtggttat tgtgtctcgt 180
catggcgttc gcgctccgac caaatttacg cagctgatgc aagatgtcac cccggacgcc 240
ttctatacgt ggccggtgaa gctgggtgaa ctgaccccgc gtggcggtga actgatcgcc 300
tatctgggtc actactggcg tcagcgcctg gtggcagatg gtctgctgcc gaaaaagggc 360
tgcccgcaga gcggtcaagt tgcaattatc gctgatgtcg acgaacgtac ccgcaaaacg 420
ggtgaagcat ttgcggccgg tctggcaccg gattgcgcca ttaccgttca tacgcaggca 480
gataccagct ctccggaccc gctgttcaac ccgctgaaaa ccggcgtctg tcagctggat 540
gtcgcgcaag tgacggacgc cattctggaa cgtgcaggcg gttccatcgc tgattttacc 600
ggtcactacc agacggcatt ccgtgaactg gaacgcgttc tgaactttcc gcagtcaaat 660
ctggcgctga aacgcgaaaa gcaggatgaa agtgcgtccc tgacccaagc cctgccgagt 720
gaactgaaag tctccgccga caatgtgtca ctgaccggcg catggtcact ggcttcgatg 780
ctgacggaaa tttttctgct gcagcaagca cagggtatgc cggaaccggg ttggggtcgt 840
atcaccgatt cgcatcagtg gaacacgctg ctgagcctgc acaatgcgca gttcgacctg 900
ctgcaacgta ccccggaagt ggcacgttcg cgcgccacgc cgctgctgga tctgattaaa 960
accgctctga cgccgcatcc gccgcagaag caagcgtatg gcgtgaccct gccgacgagc 1020
gttctgttta tcgcgggtca cgacaccaac ctggcaaatc tgggcggtgc tctggaactg 1080
cagtggaccc tgccgggtca accggataac acgccgccgg gcggtgaact ggttttcgaa 1140
cgttggcgtc gcctgagcga caattctcag tggatccaag ttagcctggt ctttcagacc 1200
ctgcagcaaa tgcgcgataa aaccccgctg ttcctgaaca cgccgccggg cgaagtgaag 1260
ctgaccctgg cgggttgcga agaacgtaac gcccagggca tgtgttctct ggcaggtttt 1320
acccagattg ttaatgaagc acgcatcccg gcttgtagtc tgtgcgtgga cggggacact 1380
ctcgtgctga caaaggagtt cgggctcatc aagatcaagg acctctacaa gattctggac 1440
ggcaagggga agaagacagt gaacggcaat gaggagtgga cagagctgga gaggccaatc 1500
actctgtacg gctacaagga cgggaagatc gtcgagatta aggctaccca cgtttacaag 1560
ggcttctccg ccgggatgat cgagattcgg acccgcacgg gccgcaagat taaggtcacg 1620
cccatccata agctcttcac aggcagggtt actaagaatg ggctggagat ccgggaggtc 1680
atggccaagg acctcaagaa gggcgatcgg atcattgtgg cgaagtaa 1728
<210> 72
<211> 575
<212> PRT
<213> Artificial Sequence
<400> 72
Met Thr His Val Leu Phe Asp Glu Ile Val Glu Ile Arg Tyr Ile Ser
1 5 10 15
Glu Gly Gln Glu Val Tyr Asp Val Thr Thr Glu Thr His Asn Phe Ile
20 25 30
Gly Gly Asn Met Pro Thr Leu Leu His Asn Ser Ala Gln Ser Glu Pro
35 40 45
Glu Leu Lys Leu Glu Ser Val Val Ile Val Ser Arg His Gly Val Arg
50 55 60
Ala Pro Thr Lys Phe Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala
65 70 75 80
Phe Tyr Thr Trp Pro Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly
85 90 95
Glu Leu Ile Ala Tyr Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala
100 105 110
Asp Gly Leu Leu Pro Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala
115 120 125
Ile Ile Ala Asp Val Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe
130 135 140
Ala Ala Gly Leu Ala Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala
145 150 155 160
Asp Thr Ser Ser Pro Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val
165 170 175
Cys Gln Leu Asp Val Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala
180 185 190
Gly Gly Ser Ile Ala Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg
195 200 205
Glu Leu Glu Arg Val Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys
210 215 220
Arg Glu Lys Gln Asp Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser
225 230 235 240
Glu Leu Lys Val Ser Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser
245 250 255
Leu Ala Ser Met Leu Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly
260 265 270
Met Pro Glu Pro Gly Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn
275 280 285
Thr Leu Leu Ser Leu His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr
290 295 300
Pro Glu Val Ala Arg Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys
305 310 315 320
Thr Ala Leu Thr Pro His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr
325 330 335
Leu Pro Thr Ser Val Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala
340 345 350
Asn Leu Gly Gly Ala Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro
355 360 365
Asp Asn Thr Pro Pro Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg
370 375 380
Leu Ser Asp Asn Ser Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr
385 390 395 400
Leu Gln Gln Met Arg Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro
405 410 415
Gly Glu Val Lys Leu Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln
420 425 430
Gly Met Cys Ser Leu Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg
435 440 445
Ile Pro Ala Cys Ser Leu Cys Val Asp Gly Asp Thr Leu Val Leu Thr
450 455 460
Lys Glu Phe Gly Leu Ile Lys Ile Lys Asp Leu Tyr Lys Ile Leu Asp
465 470 475 480
Gly Lys Gly Lys Lys Thr Val Asn Gly Asn Glu Glu Trp Thr Glu Leu
485 490 495
Glu Arg Pro Ile Thr Leu Tyr Gly Tyr Lys Asp Gly Lys Ile Val Glu
500 505 510
Ile Lys Ala Thr His Val Tyr Lys Gly Phe Ser Ala Gly Met Ile Glu
515 520 525
Ile Arg Thr Arg Thr Gly Arg Lys Ile Lys Val Thr Pro Ile His Lys
530 535 540
Leu Phe Thr Gly Arg Val Thr Lys Asn Gly Leu Glu Ile Arg Glu Val
545 550 555 560
Met Ala Lys Asp Leu Lys Lys Gly Asp Arg Ile Ile Val Ala Lys
565 570 575
<210> 73
<211> 1689
<212> DNA
<213> Artificial Sequence
<400> 73
atgaacttcg ttttcctgcc ggtggagaag atcgaggagt tcgagtacga tggctacgtc 60
tacgacgtta ctacagagac tcataatttc attgctaatg gcatcctcgt tcataatagc 120
gcccaatcgg aaccggaact gaaactggaa agtgtggtta ttgtgtctcg tcatggcgtt 180
cgcgctccga ccaaatttac gcagctgatg caagatgtca ccccggacgc cttctatacg 240
tggccggtga agctgggtga actgaccccg cgtggcggtg aactgatcgc ctatctgggt 300
cactactggc gtcagcgcct ggtggcagat ggtctgctgc cgaaaaaggg ctgcccgcag 360
agcggtcaag ttgcaattat cgctgatgtc gacgaacgta cccgcaaaac gggtgaagca 420
tttgcggccg gtctggcacc ggattgcgcc attaccgttc atacgcaggc agataccagc 480
tctccggacc cgctgttcaa cccgctgaaa accggcgtct gtcagctgga tgtcgcgcaa 540
gtgacggacg ccattctgga acgtgcaggc ggttccatcg ctgattttac cggtcactac 600
cagacggcat tccgtgaact ggaacgcgtt ctgaactttc cgcagtcaaa tctggcgctg 660
aaacgcgaaa agcaggatga aagtgcgtcc ctgacccaag ccctgccgag tgaactgaaa 720
gtctccgccg acaatgtgtc actgaccggc gcatggtcac tggcttcgat gctgacggaa 780
atttttctgc tgcagcaagc acagggtatg ccggaaccgg gttggggtcg tatcaccgat 840
tcgcatcagt ggaacacgct gctgagcctg cacaatgcgc agttcgacct gctgcaacgt 900
accccggaag tggcacgttc gcgcgccacg ccgctgctgg atctgattaa aaccgctctg 960
acgccgcatc cgccgcagaa gcaagcgtat ggcgtgaccc tgccgacgag cgttctgttt 1020
atcgcgggtc acgacaccaa cctggcaaat ctgggcggtg ctctggaact gcagtggacc 1080
ctgccgggtc aaccggataa cacgccgccg ggcggtgaac tggttttcga acgttggcgt 1140
cgcctgagcg acaattctca gtggatccaa gttagcctgg tctttcagac cctgcagcaa 1200
atgcgcgata aaaccccgct gttcctgaac acgccgccgg gcgaagtgaa gctgaccctg 1260
gcgggttgcg aagaacgtaa cgcccagggc atgtgttctc tggcaggttt tacccagatt 1320
gttaatgaag cacgcatccc ggcttgtagt ctgtgcctgc tgccggagga gcgggttatt 1380
ctgcctgact acgggcctat tactctggag gagctcttca atatgacaaa ggagacagtg 1440
ttcaaggacg aggagaagga ggtccggaag ctcggcatcc gcatgccagt ggctggcgtc 1500
gatgggcggg tgcgcctgct ggagggcccc tacgtttgga aggtgcgcta caaggggaag 1560
atgctcaggg tcaagctgaa ggactggcac agcgtggctg tcacaccaga gcatcccttc 1620
ctcaccacgc ggggctgggt gcgcgctgac cagctgaagc ccggggatta cgttgccgtg 1680
ccaaagtaa 1689
<210> 74
<211> 562
<212> PRT
<213> Artificial Sequence
<400> 74
Met Asn Phe Val Phe Leu Pro Val Glu Lys Ile Glu Glu Phe Glu Tyr
1 5 10 15
Asp Gly Tyr Val Tyr Asp Val Thr Thr Glu Thr His Asn Phe Ile Ala
20 25 30
Asn Gly Ile Leu Val His Asn Ser Ala Gln Ser Glu Pro Glu Leu Lys
35 40 45
Leu Glu Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr
50 55 60
Lys Phe Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr
65 70 75 80
Trp Pro Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile
85 90 95
Ala Tyr Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu
100 105 110
Leu Pro Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala
115 120 125
Asp Val Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly
130 135 140
Leu Ala Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser
145 150 155 160
Ser Pro Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu
165 170 175
Asp Val Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser
180 185 190
Ile Ala Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu
195 200 205
Arg Val Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys
210 215 220
Gln Asp Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys
225 230 235 240
Val Ser Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser
245 250 255
Met Leu Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu
260 265 270
Pro Gly Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu
275 280 285
Ser Leu His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val
290 295 300
Ala Arg Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu
305 310 315 320
Thr Pro His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr
325 330 335
Ser Val Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly
340 345 350
Gly Ala Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr
355 360 365
Pro Pro Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp
370 375 380
Asn Ser Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln
385 390 395 400
Met Arg Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val
405 410 415
Lys Leu Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys
420 425 430
Ser Leu Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala
435 440 445
Cys Ser Leu Cys Leu Leu Pro Glu Glu Arg Val Ile Leu Pro Asp Tyr
450 455 460
Gly Pro Ile Thr Leu Glu Glu Leu Phe Asn Met Thr Lys Glu Thr Val
465 470 475 480
Phe Lys Asp Glu Glu Lys Glu Val Arg Lys Leu Gly Ile Arg Met Pro
485 490 495
Val Ala Gly Val Asp Gly Arg Val Arg Leu Leu Glu Gly Pro Tyr Val
500 505 510
Trp Lys Val Arg Tyr Lys Gly Lys Met Leu Arg Val Lys Leu Lys Asp
515 520 525
Trp His Ser Val Ala Val Thr Pro Glu His Pro Phe Leu Thr Thr Arg
530 535 540
Gly Trp Val Arg Ala Asp Gln Leu Lys Pro Gly Asp Tyr Val Ala Val
545 550 555 560
Pro Lys
<210> 75
<211> 1728
<212> DNA
<213> Artificial Sequence
<400> 75
atgcagcata tcattttcga cgaggtcatc gatgtcaggt acattccgga gccccaggag 60
gtgtacgatg ttactacaga gactcataat ttcgtggggg gcaacatgcc aactctgctc 120
cacaatagcg cccaatcgga accggaactg aaactggaaa gtgtggttat tgtgtctcgt 180
catggcgttc gcgctccgac caaatttacg cagctgatgc aagatgtcac cccggacgcc 240
ttctatacgt ggccggtgaa gctgggtgaa ctgaccccgc gtggcggtga actgatcgcc 300
tatctgggtc actactggcg tcagcgcctg gtggcagatg gtctgctgcc gaaaaagggc 360
tgcccgcaga gcggtcaagt tgcaattatc gctgatgtcg acgaacgtac ccgcaaaacg 420
ggtgaagcat ttgcggccgg tctggcaccg gattgcgcca ttaccgttca tacgcaggca 480
gataccagct ctccggaccc gctgttcaac ccgctgaaaa ccggcgtctg tcagctggat 540
gtcgcgcaag tgacggacgc cattctggaa cgtgcaggcg gttccatcgc tgattttacc 600
ggtcactacc agacggcatt ccgtgaactg gaacgcgttc tgaactttcc gcagtcaaat 660
ctggcgctga aacgcgaaaa gcaggatgaa agtgcgtccc tgacccaagc cctgccgagt 720
gaactgaaag tctccgccga caatgtgtca ctgaccggcg catggtcact ggcttcgatg 780
ctgacggaaa tttttctgct gcagcaagca cagggtatgc cggaaccggg ttggggtcgt 840
atcaccgatt cgcatcagtg gaacacgctg ctgagcctgc acaatgcgca gttcgacctg 900
ctgcaacgta ccccggaagt ggcacgttcg cgcgccacgc cgctgctgga tctgattaaa 960
accgctctga cgccgcatcc gccgcagaag caagcgtatg gcgtgaccct gccgacgagc 1020
gttctgttta tcgcgggtca cgacaccaac ctggcaaatc tgggcggtgc tctggaactg 1080
cagtggaccc tgccgggtca accggataac acgccgccgg gcggtgaact ggttttcgaa 1140
cgttggcgtc gcctgagcga caattctcag tggatccaag ttagcctggt ctttcagacc 1200
ctgcagcaaa tgcgcgataa aaccccgctg ttcctgaaca cgccgccggg cgaagtgaag 1260
ctgaccctgg cgggttgcga agaacgtaac gcccagggca tgtgttctct ggcaggtttt 1320
acccagattg ttaatgaagc acgcatcccg gcttgtagtc tgtgcgtgga cggggacaca 1380
ctggtgctga caaaggagtt cgggctcatc aagatcaagg agctctacga gaagctggac 1440
ggcaaggggc gcaagattgt ggagggcaac gaggagtgga ccgagctgga gaagccaatc 1500
acggtctacg gctacaagga cgggaagatc gttgagatta aggccaccca cgtttacaag 1560
ggcgtgtcca gcgggatggt cgagatcagg acccggacgg gccggaagat caaggtgacg 1620
ccgattcacc gcctgttcac aggcagggtc actaaggacg ggctgatcct caaggaggtc 1680
atggctatgc atgttaagcc cggcgatagg atcgccgtgg tcaagtaa 1728
<210> 76
<211> 575
<212> PRT
<213> Artificial Sequence
<400> 76
Met Gln His Ile Ile Phe Asp Glu Val Ile Asp Val Arg Tyr Ile Pro
1 5 10 15
Glu Pro Gln Glu Val Tyr Asp Val Thr Thr Glu Thr His Asn Phe Val
20 25 30
Gly Gly Asn Met Pro Thr Leu Leu His Asn Ser Ala Gln Ser Glu Pro
35 40 45
Glu Leu Lys Leu Glu Ser Val Val Ile Val Ser Arg His Gly Val Arg
50 55 60
Ala Pro Thr Lys Phe Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala
65 70 75 80
Phe Tyr Thr Trp Pro Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly
85 90 95
Glu Leu Ile Ala Tyr Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala
100 105 110
Asp Gly Leu Leu Pro Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala
115 120 125
Ile Ile Ala Asp Val Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe
130 135 140
Ala Ala Gly Leu Ala Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala
145 150 155 160
Asp Thr Ser Ser Pro Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val
165 170 175
Cys Gln Leu Asp Val Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala
180 185 190
Gly Gly Ser Ile Ala Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg
195 200 205
Glu Leu Glu Arg Val Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys
210 215 220
Arg Glu Lys Gln Asp Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser
225 230 235 240
Glu Leu Lys Val Ser Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser
245 250 255
Leu Ala Ser Met Leu Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly
260 265 270
Met Pro Glu Pro Gly Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn
275 280 285
Thr Leu Leu Ser Leu His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr
290 295 300
Pro Glu Val Ala Arg Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys
305 310 315 320
Thr Ala Leu Thr Pro His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr
325 330 335
Leu Pro Thr Ser Val Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala
340 345 350
Asn Leu Gly Gly Ala Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro
355 360 365
Asp Asn Thr Pro Pro Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg
370 375 380
Leu Ser Asp Asn Ser Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr
385 390 395 400
Leu Gln Gln Met Arg Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro
405 410 415
Gly Glu Val Lys Leu Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln
420 425 430
Gly Met Cys Ser Leu Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg
435 440 445
Ile Pro Ala Cys Ser Leu Cys Val Asp Gly Asp Thr Leu Val Leu Thr
450 455 460
Lys Glu Phe Gly Leu Ile Lys Ile Lys Glu Leu Tyr Glu Lys Leu Asp
465 470 475 480
Gly Lys Gly Arg Lys Ile Val Glu Gly Asn Glu Glu Trp Thr Glu Leu
485 490 495
Glu Lys Pro Ile Thr Val Tyr Gly Tyr Lys Asp Gly Lys Ile Val Glu
500 505 510
Ile Lys Ala Thr His Val Tyr Lys Gly Val Ser Ser Gly Met Val Glu
515 520 525
Ile Arg Thr Arg Thr Gly Arg Lys Ile Lys Val Thr Pro Ile His Arg
530 535 540
Leu Phe Thr Gly Arg Val Thr Lys Asp Gly Leu Ile Leu Lys Glu Val
545 550 555 560
Met Ala Met His Val Lys Pro Gly Asp Arg Ile Ala Val Val Lys
565 570 575
<210> 77
<211> 1647
<212> DNA
<213> Artificial Sequence
<400> 77
atgtcagacg tctactggga tccgatcgtt tccattgagc ccgacggcgt tgaggaggtg 60
ttcgatctca ctgttccagg gccacataac ttcgttgcta atgacatcat tgctcataat 120
agcgcccaat cggaaccgga actgaaactg gaaagtgtgg ttattgtgtc tcgtcatggc 180
gttcgcgctc cgaccaaatt tacgcagctg atgcaagatg tcaccccgga cgccttctat 240
acgtggccgg tgaagctggg tgaactgacc ccgcgtggcg gtgaactgat cgcctatctg 300
ggtcactact ggcgtcagcg cctggtggca gatggtctgc tgccgaaaaa gggctgcccg 360
cagagcggtc aagttgcaat tatcgctgat gtcgacgaac gtacccgcaa aacgggtgaa 420
gcatttgcgg ccggtctggc accggattgc gccattaccg ttcatacgca ggcagatacc 480
agctctccgg acccgctgtt caacccgctg aaaaccggcg tctgtcagct ggatgtcgcg 540
caagtgacgg acgccattct ggaacgtgca ggcggttcca tcgctgattt taccggtcac 600
taccagacgg cattccgtga actggaacgc gttctgaact ttccgcagtc aaatctggcg 660
ctgaaacgcg aaaagcagga tgaaagtgcg tccctgaccc aagccctgcc gagtgaactg 720
aaagtctccg ccgacaatgt gtcactgacc ggcgcatggt cactggcttc gatgctgacg 780
gaaatttttc tgctgcagca agcacagggt atgccggaac cgggttgggg tcgtatcacc 840
gattcgcatc agtggaacac gctgctgagc ctgcacaatg cgcagttcga cctgctgcaa 900
cgtaccccgg aagtggcacg ttcgcgcgcc acgccgctgc tggatctgat taaaaccgct 960
ctgacgccgc atccgccgca gaagcaagcg tatggcgtga ccctgccgac gagcgttctg 1020
tttatcgcgg gtcacgacac caacctggca aatctgggcg gtgctctgga actgcagtgg 1080
accctgccgg gtcaaccgga taacacgccg ccgggcggtg aactggtttt cgaacgttgg 1140
cgtcgcctga gcgacaattc tcagtggatc caagttagcc tggtctttca gaccctgcag 1200
caaatgcgcg ataaaacccc gctgttcctg aacacgccgc cgggcgaagt gaagctgacc 1260
ctggcgggtt gcgaagaacg taacgcccag ggcatgtgtt ctctggcagg ttttacccag 1320
attgttaatg aagcacgcat cccggcttgt agtctgtgcc tcgcggggga cactctcatt 1380
acactggctg acgggcggcg ggttcctatt cgggagctgg tctcgcagca gaatttctcg 1440
gtctgggcgc tgaacccgca gacgtacagg ctggagaggg ctcgggtctc ccgggccttc 1500
tgcacaggca tcaagcccgt ttacaggctg accacgaggc tcgggaggag cattagggct 1560
actgctaatc accgcttcct gaccccacag ggctggaaga gggtggacga gctccagcct 1620
ggggattacc tggctctccc aaggtaa 1647
<210> 78
<211> 548
<212> PRT
<213> Artificial Sequence
<400> 78
Met Ser Asp Val Tyr Trp Asp Pro Ile Val Ser Ile Glu Pro Asp Gly
1 5 10 15
Val Glu Glu Val Phe Asp Leu Thr Val Pro Gly Pro His Asn Phe Val
20 25 30
Ala Asn Asp Ile Ile Ala His Asn Ser Ala Gln Ser Glu Pro Glu Leu
35 40 45
Lys Leu Glu Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala Pro
50 55 60
Thr Lys Phe Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr
65 70 75 80
Thr Trp Pro Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu
85 90 95
Ile Ala Tyr Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly
100 105 110
Leu Leu Pro Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile
115 120 125
Ala Asp Val Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala
130 135 140
Gly Leu Ala Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr
145 150 155 160
Ser Ser Pro Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln
165 170 175
Leu Asp Val Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly
180 185 190
Ser Ile Ala Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu
195 200 205
Glu Arg Val Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu
210 215 220
Lys Gln Asp Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu
225 230 235 240
Lys Val Ser Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala
245 250 255
Ser Met Leu Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro
260 265 270
Glu Pro Gly Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu
275 280 285
Leu Ser Leu His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu
290 295 300
Val Ala Arg Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala
305 310 315 320
Leu Thr Pro His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro
325 330 335
Thr Ser Val Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu
340 345 350
Gly Gly Ala Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn
355 360 365
Thr Pro Pro Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser
370 375 380
Asp Asn Ser Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln
385 390 395 400
Gln Met Arg Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu
405 410 415
Val Lys Leu Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met
420 425 430
Cys Ser Leu Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro
435 440 445
Ala Cys Ser Leu Cys Leu Ala Gly Asp Thr Leu Ile Thr Leu Ala Asp
450 455 460
Gly Arg Arg Val Pro Ile Arg Glu Leu Val Ser Gln Gln Asn Phe Ser
465 470 475 480
Val Trp Ala Leu Asn Pro Gln Thr Tyr Arg Leu Glu Arg Ala Arg Val
485 490 495
Ser Arg Ala Phe Cys Thr Gly Ile Lys Pro Val Tyr Arg Leu Thr Thr
500 505 510
Arg Leu Gly Arg Ser Ile Arg Ala Thr Ala Asn His Arg Phe Leu Thr
515 520 525
Pro Gln Gly Trp Lys Arg Val Asp Glu Leu Gln Pro Gly Asp Tyr Leu
530 535 540
Ala Leu Pro Arg
545
<210> 79
<211> 1620
<212> DNA
<213> Artificial Sequence
<400> 79
atgtggcgga tgaccggcat cgatgtcgag cccgacggcg ttggggatta cttcggcttc 60
actctggatg gcaatgggcg cttcctcctc ggggatggca ctgttactca taatagcgcc 120
caatcggaac cggaactgaa actggaaagt gtggttattg tgtctcgtca tggcgttcgc 180
gctccgacca aatttacgca gctgatgcaa gatgtcaccc cggacgcctt ctatacgtgg 240
ccggtgaagc tgggtgaact gaccccgcgt ggcggtgaac tgatcgccta tctgggtcac 300
tactggcgtc agcgcctggt ggcagatggt ctgctgccga aaaagggctg cccgcagagc 360
ggtcaagttg caattatcgc tgatgtcgac gaacgtaccc gcaaaacggg tgaagcattt 420
gcggccggtc tggcaccgga ttgcgccatt accgttcata cgcaggcaga taccagctct 480
ccggacccgc tgttcaaccc gctgaaaacc ggcgtctgtc agctggatgt cgcgcaagtg 540
acggacgcca ttctggaacg tgcaggcggt tccatcgctg attttaccgg tcactaccag 600
acggcattcc gtgaactgga acgcgttctg aactttccgc agtcaaatct ggcgctgaaa 660
cgcgaaaagc aggatgaaag tgcgtccctg acccaagccc tgccgagtga actgaaagtc 720
tccgccgaca atgtgtcact gaccggcgca tggtcactgg cttcgatgct gacggaaatt 780
tttctgctgc agcaagcaca gggtatgccg gaaccgggtt ggggtcgtat caccgattcg 840
catcagtgga acacgctgct gagcctgcac aatgcgcagt tcgacctgct gcaacgtacc 900
ccggaagtgg cacgttcgcg cgccacgccg ctgctggatc tgattaaaac cgctctgacg 960
ccgcatccgc cgcagaagca agcgtatggc gtgaccctgc cgacgagcgt tctgtttatc 1020
gcgggtcacg acaccaacct ggcaaatctg ggcggtgctc tggaactgca gtggaccctg 1080
ccgggtcaac cggataacac gccgccgggc ggtgaactgg ttttcgaacg ttggcgtcgc 1140
ctgagcgaca attctcagtg gatccaagtt agcctggtct ttcagaccct gcagcaaatg 1200
cgcgataaaa ccccgctgtt cctgaacacg ccgccgggcg aagtgaagct gaccctggcg 1260
ggttgcgaag aacgtaacgc ccagggcatg tgttctctgg caggttttac ccagattgtt 1320
aatgaagcac gcatcccggc ttgtagtctg tgcctcggga aggggacacc ggttatgatg 1380
tacgatgggc ggacaaagcc agtggagaag gtggaggtcg gggacaggct catgggggac 1440
gatggcagcc caaggacggt gcagtcgctg gccaggggga gggagcagat gtactgggtc 1500
cgccagaaga ggggcatgga ctacagggtt aacgagagcc acatcctctc gctgaagaag 1560
tctaggaggg agggcgcccg cgacaggggg tcaatcgcgg atatttccgt ccgcgactaa 1620
<210> 80
<211> 539
<212> PRT
<213> Artificial Sequence
<400> 80
Met Trp Arg Met Thr Gly Ile Asp Val Glu Pro Asp Gly Val Gly Asp
1 5 10 15
Tyr Phe Gly Phe Thr Leu Asp Gly Asn Gly Arg Phe Leu Leu Gly Asp
20 25 30
Gly Thr Val Thr His Asn Ser Ala Gln Ser Glu Pro Glu Leu Lys Leu
35 40 45
Glu Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr Lys
50 55 60
Phe Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp
65 70 75 80
Pro Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala
85 90 95
Tyr Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu
100 105 110
Pro Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp
115 120 125
Val Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu
130 135 140
Ala Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser
145 150 155 160
Pro Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp
165 170 175
Val Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile
180 185 190
Ala Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg
195 200 205
Val Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln
210 215 220
Asp Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val
225 230 235 240
Ser Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met
245 250 255
Leu Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro
260 265 270
Gly Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser
275 280 285
Leu His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala
290 295 300
Arg Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr
305 310 315 320
Pro His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser
325 330 335
Val Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly
340 345 350
Ala Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro
355 360 365
Pro Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn
370 375 380
Ser Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met
385 390 395 400
Arg Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys
405 410 415
Leu Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser
420 425 430
Leu Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys
435 440 445
Ser Leu Cys Leu Gly Lys Gly Thr Pro Val Met Met Tyr Asp Gly Arg
450 455 460
Thr Lys Pro Val Glu Lys Val Glu Val Gly Asp Arg Leu Met Gly Asp
465 470 475 480
Asp Gly Ser Pro Arg Thr Val Gln Ser Leu Ala Arg Gly Arg Glu Gln
485 490 495
Met Tyr Trp Val Arg Gln Lys Arg Gly Met Asp Tyr Arg Val Asn Glu
500 505 510
Ser His Ile Leu Ser Leu Lys Lys Ser Arg Arg Glu Gly Ala Arg Asp
515 520 525
Arg Gly Ser Ile Ala Asp Ile Ser Val Arg Asp
530 535
<210> 81
<211> 1647
<212> DNA
<213> Artificial Sequence
<400> 81
atgaattctt tctacaatct gtcaaccttc gaggtgtcat ccgagtacta caagggcgag 60
gtctacgatc tcactctgga gggcaatcct tactacttcg ccaatggcat cctcacacat 120
aatagcgccc aatcggaacc ggaactgaaa ctggaaagtg tggttattgt gtctcgtcat 180
ggcgttcgcg ctccgaccaa atttacgcag ctgatgcaag atgtcacccc ggacgccttc 240
tatacgtggc cggtgaagct gggtgaactg accccgcgtg gcggtgaact gatcgcctat 300
ctgggtcact actggcgtca gcgcctggtg gcagatggtc tgctgccgaa aaagggctgc 360
ccgcagagcg gtcaagttgc aattatcgct gatgtcgacg aacgtacccg caaaacgggt 420
gaagcatttg cggccggtct ggcaccggat tgcgccatta ccgttcatac gcaggcagat 480
accagctctc cggacccgct gttcaacccg ctgaaaaccg gcgtctgtca gctggatgtc 540
gcgcaagtga cggacgccat tctggaacgt gcaggcggtt ccatcgctga ttttaccggt 600
cactaccaga cggcattccg tgaactggaa cgcgttctga actttccgca gtcaaatctg 660
gcgctgaaac gcgaaaagca ggatgaaagt gcgtccctga cccaagccct gccgagtgaa 720
ctgaaagtct ccgccgacaa tgtgtcactg accggcgcat ggtcactggc ttcgatgctg 780
acggaaattt ttctgctgca gcaagcacag ggtatgccgg aaccgggttg gggtcgtatc 840
accgattcgc atcagtggaa cacgctgctg agcctgcaca atgcgcagtt cgacctgctg 900
caacgtaccc cggaagtggc acgttcgcgc gccacgccgc tgctggatct gattaaaacc 960
gctctgacgc cgcatccgcc gcagaagcaa gcgtatggcg tgaccctgcc gacgagcgtt 1020
ctgtttatcg cgggtcacga caccaacctg gcaaatctgg gcggtgctct ggaactgcag 1080
tggaccctgc cgggtcaacc ggataacacg ccgccgggcg gtgaactggt tttcgaacgt 1140
tggcgtcgcc tgagcgacaa ttctcagtgg atccaagtta gcctggtctt tcagaccctg 1200
cagcaaatgc gcgataaaac cccgctgttc ctgaacacgc cgccgggcga agtgaagctg 1260
accctggcgg gttgcgaaga acgtaacgcc cagggcatgt gttctctggc aggttttacc 1320
cagattgtta atgaagcacg catcccggct tgtagtctgt gccatcctgc ggacactaag 1380
gtcatcgtga agggcaaggg catcgttaat atctcggacg tgaaggaggg ggactacatt 1440
ctcggcatcg acggctggca gcgggtcaag aaggtttgga agtaccacta cgagggcaag 1500
ctcatcaaca ttaatgggct gaagtgcacg ccgaaccata aggttcccgt ggtcacagag 1560
aatgacaggc agactcgcat cagggattcc ctcgccaaga gcttcctgtc gggcaaggtc 1620
aaggggaaga tcattaccac gaagtaa 1647
<210> 82
<211> 548
<212> PRT
<213> Artificial Sequence
<400> 82
Met Asn Ser Phe Tyr Asn Leu Ser Thr Phe Glu Val Ser Ser Glu Tyr
1 5 10 15
Tyr Lys Gly Glu Val Tyr Asp Leu Thr Leu Glu Gly Asn Pro Tyr Tyr
20 25 30
Phe Ala Asn Gly Ile Leu Thr His Asn Ser Ala Gln Ser Glu Pro Glu
35 40 45
Leu Lys Leu Glu Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala
50 55 60
Pro Thr Lys Phe Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe
65 70 75 80
Tyr Thr Trp Pro Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu
85 90 95
Leu Ile Ala Tyr Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp
100 105 110
Gly Leu Leu Pro Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile
115 120 125
Ile Ala Asp Val Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala
130 135 140
Ala Gly Leu Ala Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp
145 150 155 160
Thr Ser Ser Pro Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys
165 170 175
Gln Leu Asp Val Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly
180 185 190
Gly Ser Ile Ala Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu
195 200 205
Leu Glu Arg Val Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg
210 215 220
Glu Lys Gln Asp Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu
225 230 235 240
Leu Lys Val Ser Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu
245 250 255
Ala Ser Met Leu Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met
260 265 270
Pro Glu Pro Gly Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr
275 280 285
Leu Leu Ser Leu His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro
290 295 300
Glu Val Ala Arg Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr
305 310 315 320
Ala Leu Thr Pro His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu
325 330 335
Pro Thr Ser Val Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn
340 345 350
Leu Gly Gly Ala Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp
355 360 365
Asn Thr Pro Pro Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu
370 375 380
Ser Asp Asn Ser Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu
385 390 395 400
Gln Gln Met Arg Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly
405 410 415
Glu Val Lys Leu Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly
420 425 430
Met Cys Ser Leu Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile
435 440 445
Pro Ala Cys Ser Leu Cys His Pro Ala Asp Thr Lys Val Ile Val Lys
450 455 460
Gly Lys Gly Ile Val Asn Ile Ser Asp Val Lys Glu Gly Asp Tyr Ile
465 470 475 480
Leu Gly Ile Asp Gly Trp Gln Arg Val Lys Lys Val Trp Lys Tyr His
485 490 495
Tyr Glu Gly Lys Leu Ile Asn Ile Asn Gly Leu Lys Cys Thr Pro Asn
500 505 510
His Lys Val Pro Val Val Thr Glu Asn Asp Arg Gln Thr Arg Ile Arg
515 520 525
Asp Ser Leu Ala Lys Ser Phe Leu Ser Gly Lys Val Lys Gly Lys Ile
530 535 540
Ile Thr Thr Lys
545
<210> 83
<211> 1707
<212> DNA
<213> Artificial Sequence
<400> 83
atgtcgaagt gcgtcctcaa ctactcgccc tacaagatcg agtctgttaa tattggcgct 60
gtgtgcgact acagctacga tttcgccatc gagggcatca atgataatga ctcttggtac 120
tggcaggggg ctctcaagtc tcacaacagc gcccaatcgg aaccggaact gaaactggaa 180
agtgtggtta ttgtgtctcg tcatggcgtt cgcgctccga ccaaatttac gcagctgatg 240
caagatgtca ccccggacgc cttctatacg tggccggtga agctgggtga actgaccccg 300
cgtggcggtg aactgatcgc ctatctgggt cactactggc gtcagcgcct ggtggcagat 360
ggtctgctgc cgaaaaaggg ctgcccgcag agcggtcaag ttgcaattat cgctgatgtc 420
gacgaacgta cccgcaaaac gggtgaagca tttgcggccg gtctggcacc ggattgcgcc 480
attaccgttc atacgcaggc agataccagc tctccggacc cgctgttcaa cccgctgaaa 540
accggcgtct gtcagctgga tgtcgcgcaa gtgacggacg ccattctgga acgtgcaggc 600
ggttccatcg ctgattttac cggtcactac cagacggcat tccgtgaact ggaacgcgtt 660
ctgaactttc cgcagtcaaa tctggcgctg aaacgcgaaa agcaggatga aagtgcgtcc 720
ctgacccaag ccctgccgag tgaactgaaa gtctccgccg acaatgtgtc actgaccggc 780
gcatggtcac tggcttcgat gctgacggaa atttttctgc tgcagcaagc acagggtatg 840
ccggaaccgg gttggggtcg tatcaccgat tcgcatcagt ggaacacgct gctgagcctg 900
cacaatgcgc agttcgacct gctgcaacgt accccggaag tggcacgttc gcgcgccacg 960
ccgctgctgg atctgattaa aaccgctctg acgccgcatc cgccgcagaa gcaagcgtat 1020
ggcgtgaccc tgccgacgag cgttctgttt atcgcgggtc acgacaccaa cctggcaaat 1080
ctgggcggtg ctctggaact gcagtggacc ctgccgggtc aaccggataa cacgccgccg 1140
ggcggtgaac tggttttcga acgttggcgt cgcctgagcg acaattctca gtggatccaa 1200
gttagcctgg tctttcagac cctgcagcaa atgcgcgata aaaccccgct gttcctgaac 1260
acgccgccgg gcgaagtgaa gctgaccctg gcgggttgcg aagaacgtaa cgcccagggc 1320
atgtgttctc tggcaggttt tacccagatt gttaatgaag cacgcatccc ggcttgtagt 1380
ctgtgcctgg acaagacggc tctgcggatt ttcaatcagg ggctgctcta cgcggatgag 1440
gtcgtgacac cgggctcggg ggagacagtc ggcctcgggc tgacggtcag gaacggcatc 1500
ggggcgtcca cagccattgc gaatcagccg atggagctgg ttgagatcaa gctcgctaac 1560
ggccggaagc tgcgcatgac ccctaatcac cggatgtccg tgaagggcaa gtggattcat 1620
gcctgcaacc tcaagccggg gatgctcctg gactacagca tcggcgagta ccagaagcgc 1680
gaggacaccc tcctgattcc tctctaa 1707
<210> 84
<211> 568
<212> PRT
<213> Artificial Sequence
<400> 84
Met Ser Lys Cys Val Leu Asn Tyr Ser Pro Tyr Lys Ile Glu Ser Val
1 5 10 15
Asn Ile Gly Ala Val Cys Asp Tyr Ser Tyr Asp Phe Ala Ile Glu Gly
20 25 30
Ile Asn Asp Asn Asp Ser Trp Tyr Trp Gln Gly Ala Leu Lys Ser His
35 40 45
Asn Ser Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val Val Ile
50 55 60
Val Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe Thr Gln Leu Met
65 70 75 80
Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro Val Lys Leu Gly
85 90 95
Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu Gly His Tyr
100 105 110
Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys Lys Gly Cys
115 120 125
Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp Glu Arg Thr
130 135 140
Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro Asp Cys Ala
145 150 155 160
Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro Asp Pro Leu Phe
165 170 175
Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val Ala Gln Val Thr
180 185 190
Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp Phe Thr Gly
195 200 205
His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu Asn Phe Pro
210 215 220
Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp Glu Ser Ala Ser
225 230 235 240
Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala Asp Asn Val
245 250 255
Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu Thr Glu Ile Phe
260 265 270
Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp Gly Arg Ile
275 280 285
Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu His Asn Ala Gln
290 295 300
Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser Arg Ala Thr
305 310 315 320
Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His Pro Pro Gln
325 330 335
Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu Phe Ile Ala
340 345 350
Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu Glu Leu Gln
355 360 365
Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly Glu Leu
370 375 380
Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln Trp Ile Gln
385 390 395 400
Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp Lys Thr Pro
405 410 415
Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr Leu Ala Gly
420 425 430
Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala Gly Phe Thr
435 440 445
Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu Cys Leu Asp
450 455 460
Lys Thr Ala Leu Arg Ile Phe Asn Gln Gly Leu Leu Tyr Ala Asp Glu
465 470 475 480
Val Val Thr Pro Gly Ser Gly Glu Thr Val Gly Leu Gly Leu Thr Val
485 490 495
Arg Asn Gly Ile Gly Ala Ser Thr Ala Ile Ala Asn Gln Pro Met Glu
500 505 510
Leu Val Glu Ile Lys Leu Ala Asn Gly Arg Lys Leu Arg Met Thr Pro
515 520 525
Asn His Arg Met Ser Val Lys Gly Lys Trp Ile His Ala Cys Asn Leu
530 535 540
Lys Pro Gly Met Leu Leu Asp Tyr Ser Ile Gly Glu Tyr Gln Lys Arg
545 550 555 560
Glu Asp Thr Leu Leu Ile Pro Leu
565
<210> 85
<211> 1689
<212> DNA
<213> Artificial Sequence
<400> 85
atgaatctcg tcttcatccc ggttgaggac attgaggagt tcgagtacga gggctacgtt 60
tacgacgtta ctacagagac tcataatttc gttgctaatg gcatcctcgt tcataatagc 120
gcccaatcgg aaccggaact gaaactggaa agtgtggtta ttgtgtctcg tcatggcgtt 180
cgcgctccga ccaaatttac gcagctgatg caagatgtca ccccggacgc cttctatacg 240
tggccggtga agctgggtga actgaccccg cgtggcggtg aactgatcgc ctatctgggt 300
cactactggc gtcagcgcct ggtggcagat ggtctgctgc cgaaaaaggg ctgcccgcag 360
agcggtcaag ttgcaattat cgctgatgtc gacgaacgta cccgcaaaac gggtgaagca 420
tttgcggccg gtctggcacc ggattgcgcc attaccgttc atacgcaggc agataccagc 480
tctccggacc cgctgttcaa cccgctgaaa accggcgtct gtcagctgga tgtcgcgcaa 540
gtgacggacg ccattctgga acgtgcaggc ggttccatcg ctgattttac cggtcactac 600
cagacggcat tccgtgaact ggaacgcgtt ctgaactttc cgcagtcaaa tctggcgctg 660
aaacgcgaaa agcaggatga aagtgcgtcc ctgacccaag ccctgccgag tgaactgaaa 720
gtctccgccg acaatgtgtc actgaccggc gcatggtcac tggcttcgat gctgacggaa 780
atttttctgc tgcagcaagc acagggtatg ccggaaccgg gttggggtcg tatcaccgat 840
tcgcatcagt ggaacacgct gctgagcctg cacaatgcgc agttcgacct gctgcaacgt 900
accccggaag tggcacgttc gcgcgccacg ccgctgctgg atctgattaa aaccgctctg 960
acgccgcatc cgccgcagaa gcaagcgtat ggcgtgaccc tgccgacgag cgttctgttt 1020
atcgcgggtc acgacaccaa cctggcaaat ctgggcggtg ctctggaact gcagtggacc 1080
ctgccgggtc aaccggataa cacgccgccg ggcggtgaac tggttttcga acgttggcgt 1140
cgcctgagcg acaattctca gtggatccaa gttagcctgg tctttcagac cctgcagcaa 1200
atgcgcgata aaaccccgct gttcctgaac acgccgccgg gcgaagtgaa gctgaccctg 1260
gcgggttgcg aagaacgtaa cgcccagggc atgtgttctc tggcaggttt tacccagatt 1320
gttaatgaag cacgcatccc ggcttgtagt ctgtgcctgc tgccggatga gaaggttatt 1380
ctccctgagc atgggcctat tacactcaag gggctcttcg atctcgctaa ggagacagtc 1440
gtggctgaca acgagaagga gatccgcaag ctgggcgcca agctcaccat tgtgggcgag 1500
gatgggaggc tcagggtcct ggagagccca tacgtttgga aggtgcggca ccgcggcaag 1560
atgctgaggg tcaagctcaa gaactggcac tcagtgtccg tcacgccaga gcatcccttc 1620
ctgaccacgc ggggctgggt gcgcgctgac cagctcaagc cgggggatta cgttgcggtg 1680
cccaggtaa 1689
<210> 86
<211> 562
<212> PRT
<213> Artificial Sequence
<400> 86
Met Asn Leu Val Phe Ile Pro Val Glu Asp Ile Glu Glu Phe Glu Tyr
1 5 10 15
Glu Gly Tyr Val Tyr Asp Val Thr Thr Glu Thr His Asn Phe Val Ala
20 25 30
Asn Gly Ile Leu Val His Asn Ser Ala Gln Ser Glu Pro Glu Leu Lys
35 40 45
Leu Glu Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr
50 55 60
Lys Phe Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr
65 70 75 80
Trp Pro Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile
85 90 95
Ala Tyr Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu
100 105 110
Leu Pro Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala
115 120 125
Asp Val Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly
130 135 140
Leu Ala Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser
145 150 155 160
Ser Pro Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu
165 170 175
Asp Val Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser
180 185 190
Ile Ala Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu
195 200 205
Arg Val Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys
210 215 220
Gln Asp Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys
225 230 235 240
Val Ser Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser
245 250 255
Met Leu Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu
260 265 270
Pro Gly Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu
275 280 285
Ser Leu His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val
290 295 300
Ala Arg Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu
305 310 315 320
Thr Pro His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr
325 330 335
Ser Val Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly
340 345 350
Gly Ala Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr
355 360 365
Pro Pro Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp
370 375 380
Asn Ser Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln
385 390 395 400
Met Arg Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val
405 410 415
Lys Leu Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys
420 425 430
Ser Leu Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala
435 440 445
Cys Ser Leu Cys Leu Leu Pro Asp Glu Lys Val Ile Leu Pro Glu His
450 455 460
Gly Pro Ile Thr Leu Lys Gly Leu Phe Asp Leu Ala Lys Glu Thr Val
465 470 475 480
Val Ala Asp Asn Glu Lys Glu Ile Arg Lys Leu Gly Ala Lys Leu Thr
485 490 495
Ile Val Gly Glu Asp Gly Arg Leu Arg Val Leu Glu Ser Pro Tyr Val
500 505 510
Trp Lys Val Arg His Arg Gly Lys Met Leu Arg Val Lys Leu Lys Asn
515 520 525
Trp His Ser Val Ser Val Thr Pro Glu His Pro Phe Leu Thr Thr Arg
530 535 540
Gly Trp Val Arg Ala Asp Gln Leu Lys Pro Gly Asp Tyr Val Ala Val
545 550 555 560
Pro Arg
<210> 87
<211> 1656
<212> DNA
<213> Artificial Sequence
<400> 87
atggctgagg tttactggga tcgcgtcgag gcggttgagc cgctcggcga ggaggaggtc 60
ttcgatctca ctgtggaggg cactcatact ttcgttgcgg aggatgttat cgttcataat 120
agcgcccaat cggaaccgga actgaaactg gaaagtgtgg ttattgtgtc tcgtcatggc 180
gttcgcgctc cgaccaaatt tacgcagctg atgcaagatg tcaccccgga cgccttctat 240
acgtggccgg tgaagctggg tgaactgacc ccgcgtggcg gtgaactgat cgcctatctg 300
ggtcactact ggcgtcagcg cctggtggca gatggtctgc tgccgaaaaa gggctgcccg 360
cagagcggtc aagttgcaat tatcgctgat gtcgacgaac gtacccgcaa aacgggtgaa 420
gcatttgcgg ccggtctggc accggattgc gccattaccg ttcatacgca ggcagatacc 480
agctctccgg acccgctgtt caacccgctg aaaaccggcg tctgtcagct ggatgtcgcg 540
caagtgacgg acgccattct ggaacgtgca ggcggttcca tcgctgattt taccggtcac 600
taccagacgg cattccgtga actggaacgc gttctgaact ttccgcagtc aaatctggcg 660
ctgaaacgcg aaaagcagga tgaaagtgcg tccctgaccc aagccctgcc gagtgaactg 720
aaagtctccg ccgacaatgt gtcactgacc ggcgcatggt cactggcttc gatgctgacg 780
gaaatttttc tgctgcagca agcacagggt atgccggaac cgggttgggg tcgtatcacc 840
gattcgcatc agtggaacac gctgctgagc ctgcacaatg cgcagttcga cctgctgcaa 900
cgtaccccgg aagtggcacg ttcgcgcgcc acgccgctgc tggatctgat taaaaccgct 960
ctgacgccgc atccgccgca gaagcaagcg tatggcgtga ccctgccgac gagcgttctg 1020
tttatcgcgg gtcacgacac caacctggca aatctgggcg gtgctctgga actgcagtgg 1080
accctgccgg gtcaaccgga taacacgccg ccgggcggtg aactggtttt cgaacgttgg 1140
cgtcgcctga gcgacaattc tcagtggatc caagttagcc tggtctttca gaccctgcag 1200
caaatgcgcg ataaaacccc gctgttcctg aacacgccgc cgggcgaagt gaagctgacc 1260
ctggcgggtt gcgaagaacg taacgcccag ggcatgtgtt ctctggcagg ttttacccag 1320
attgttaatg aagcacgcat cccggcttgt agtctgtgcc tgcctgcgcg ggctagggtc 1380
gtggattggt gcacagggcg ggtcgttcgg gtcggggaga tcgttagggg ggaggctaag 1440
ggcgtctggg tggtctccct ggacgaggct aggctgaggc tcgttccaag gcctgttgtg 1500
gctgctttcc caagcggcaa ggctcaggtg tacgctctga ggaccgctac gggcagggtg 1560
ctggaggcga cagctaacca cccagtctac actccagagg gctggaggcc actggggacc 1620
ctcgctcctg gcgactacgt cgctctgcca aggtaa 1656
<210> 88
<211> 551
<212> PRT
<213> Artificial Sequence
<400> 88
Met Ala Glu Val Tyr Trp Asp Arg Val Glu Ala Val Glu Pro Leu Gly
1 5 10 15
Glu Glu Glu Val Phe Asp Leu Thr Val Glu Gly Thr His Thr Phe Val
20 25 30
Ala Glu Asp Val Ile Val His Asn Ser Ala Gln Ser Glu Pro Glu Leu
35 40 45
Lys Leu Glu Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala Pro
50 55 60
Thr Lys Phe Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr
65 70 75 80
Thr Trp Pro Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu
85 90 95
Ile Ala Tyr Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly
100 105 110
Leu Leu Pro Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile
115 120 125
Ala Asp Val Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala
130 135 140
Gly Leu Ala Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr
145 150 155 160
Ser Ser Pro Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln
165 170 175
Leu Asp Val Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly
180 185 190
Ser Ile Ala Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu
195 200 205
Glu Arg Val Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu
210 215 220
Lys Gln Asp Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu
225 230 235 240
Lys Val Ser Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala
245 250 255
Ser Met Leu Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro
260 265 270
Glu Pro Gly Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu
275 280 285
Leu Ser Leu His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu
290 295 300
Val Ala Arg Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala
305 310 315 320
Leu Thr Pro His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro
325 330 335
Thr Ser Val Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu
340 345 350
Gly Gly Ala Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn
355 360 365
Thr Pro Pro Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser
370 375 380
Asp Asn Ser Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln
385 390 395 400
Gln Met Arg Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu
405 410 415
Val Lys Leu Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met
420 425 430
Cys Ser Leu Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro
435 440 445
Ala Cys Ser Leu Cys Leu Pro Ala Arg Ala Arg Val Val Asp Trp Cys
450 455 460
Thr Gly Arg Val Val Arg Val Gly Glu Ile Val Arg Gly Glu Ala Lys
465 470 475 480
Gly Val Trp Val Val Ser Leu Asp Glu Ala Arg Leu Arg Leu Val Pro
485 490 495
Arg Pro Val Val Ala Ala Phe Pro Ser Gly Lys Ala Gln Val Tyr Ala
500 505 510
Leu Arg Thr Ala Thr Gly Arg Val Leu Glu Ala Thr Ala Asn His Pro
515 520 525
Val Tyr Thr Pro Glu Gly Trp Arg Pro Leu Gly Thr Leu Ala Pro Gly
530 535 540
Asp Tyr Val Ala Leu Pro Arg
545 550
<210> 89
<211> 1716
<212> DNA
<213> Artificial Sequence
<400> 89
atggttaagg tgattggaag acgttctctt ggtgttcaaa ggatcttcga tatcggattg 60
ccacaagacc acaactttct tctcgctaat ggtgccatcg ctgccaatag cgcccaatcg 120
gaaccggaac tgaaactgga aagtgtggtt attgtgtctc gtcatggcgt tcgcgctccg 180
accaaattta cgcagctgat gcaagatgtc accccggacg ccttctatac gtggccggtg 240
aagctgggtg aactgacccc gcgtggcggt gaactgatcg cctatctggg tcactactgg 300
cgtcagcgcc tggtggcaga tggtctgctg ccgaaaaagg gctgcccgca gagcggtcaa 360
gttgcaatta tcgctgatgt cgacgaacgt acccgcaaaa cgggtgaagc atttgcggcc 420
ggtctggcac cggattgcgc cattaccgtt catacgcagg cagataccag ctctccggac 480
ccgctgttca acccgctgaa aaccggcgtc tgtcagctgg atgtcgcgca agtgacggac 540
gccattctgg aacgtgcagg cggttccatc gctgatttta ccggtcacta ccagacggca 600
ttccgtgaac tggaacgcgt tctgaacttt ccgcagtcaa atctggcgct gaaacgcgaa 660
aagcaggatg aaagtgcgtc cctgacccaa gccctgccga gtgaactgaa agtctccgcc 720
gacaatgtgt cactgaccgg cgcatggtca ctggcttcga tgctgacgga aatttttctg 780
ctgcagcaag cacagggtat gccggaaccg ggttggggtc gtatcaccga ttcgcatcag 840
tggaacacgc tgctgagcct gcacaatgcg cagttcgacc tgctgcaacg taccccggaa 900
gtggcacgtt cgcgcgccac gccgctgctg gatctgatta aaaccgctct gacgccgcat 960
ccgccgcaga agcaagcgta tggcgtgacc ctgccgacga gcgttctgtt tatcgcgggt 1020
cacgacacca acctggcaaa tctgggcggt gctctggaac tgcagtggac cctgccgggt 1080
caaccggata acacgccgcc gggcggtgaa ctggttttcg aacgttggcg tcgcctgagc 1140
gacaattctc agtggatcca agttagcctg gtctttcaga ccctgcagca aatgcgcgat 1200
aaaaccccgc tgttcctgaa cacgccgccg ggcgaagtga agctgaccct ggcgggttgc 1260
gaagaacgta acgcccaggg catgtgttct ctggcaggtt ttacccagat tgttaatgaa 1320
gcacgcatcc cggcttgtag tctgtgcctt tctttcggaa ctgagatcct taccgttgag 1380
tacggaccac ttcctattgg taagatcgtt tctgaggaaa ttaactgctc agtgtactct 1440
gttgatccag aaggaagagt ttacactcag gctatcgcac aatggcacga taggggtgaa 1500
caagaggttc tggagtacga gcttgaagat ggatccgtta ttcgtgctac ctctgaccat 1560
agattcttga ctacagatta tcagcttctc gctatcgagg aaatctttgc taggcaactt 1620
gatctcctta ctttggagaa catcaagcag acagaagagg ctcttgacaa ccacagactt 1680
ccattccctt tgctcgatgc tggaaccatc aagtaa 1716
<210> 90
<211> 571
<212> PRT
<213> Artificial Sequence
<400> 90
Met Val Lys Val Ile Gly Arg Arg Ser Leu Gly Val Gln Arg Ile Phe
1 5 10 15
Asp Ile Gly Leu Pro Gln Asp His Asn Phe Leu Leu Ala Asn Gly Ala
20 25 30
Ile Ala Ala Asn Ser Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser
35 40 45
Val Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe Thr
50 55 60
Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro Val
65 70 75 80
Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu
85 90 95
Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys
100 105 110
Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp
115 120 125
Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro
130 135 140
Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro Asp
145 150 155 160
Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val Ala
165 170 175
Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp
180 185 190
Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu
195 200 205
Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp Glu
210 215 220
Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala
225 230 235 240
Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu Thr
245 250 255
Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp
260 265 270
Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu His
275 280 285
Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser
290 295 300
Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His
305 310 315 320
Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu
325 330 335
Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu
340 345 350
Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly
355 360 365
Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln
370 375 380
Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp
385 390 395 400
Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr
405 410 415
Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala
420 425 430
Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu
435 440 445
Cys Leu Ser Phe Gly Thr Glu Ile Leu Thr Val Glu Tyr Gly Pro Leu
450 455 460
Pro Ile Gly Lys Ile Val Ser Glu Glu Ile Asn Cys Ser Val Tyr Ser
465 470 475 480
Val Asp Pro Glu Gly Arg Val Tyr Thr Gln Ala Ile Ala Gln Trp His
485 490 495
Asp Arg Gly Glu Gln Glu Val Leu Glu Tyr Glu Leu Glu Asp Gly Ser
500 505 510
Val Ile Arg Ala Thr Ser Asp His Arg Phe Leu Thr Thr Asp Tyr Gln
515 520 525
Leu Leu Ala Ile Glu Glu Ile Phe Ala Arg Gln Leu Asp Leu Leu Thr
530 535 540
Leu Glu Asn Ile Lys Gln Thr Glu Glu Ala Leu Asp Asn His Arg Leu
545 550 555 560
Pro Phe Pro Leu Leu Asp Ala Gly Thr Ile Lys
565 570
<210> 91
<211> 1614
<212> DNA
<213> Artificial Sequence
<400> 91
atgatgctga agaaaattct gaagatcgaa gaactggatg aacgtgaact gattgacatc 60
gaagttagcg gcaaccatct gttttacgcg aatgacattc tgacccacaa cagcgcccaa 120
tcggaaccgg aactgaaact ggaaagtgtg gttattgtgt ctcgtcatgg cgttcgcgct 180
ccgaccaaat ttacgcagct gatgcaagat gtcaccccgg acgccttcta tacgtggccg 240
gtgaagctgg gtgaactgac cccgcgtggc ggtgaactga tcgcctatct gggtcactac 300
tggcgtcagc gcctggtggc agatggtctg ctgccgaaaa agggctgccc gcagagcggt 360
caagttgcaa ttatcgctga tgtcgacgaa cgtacccgca aaacgggtga agcatttgcg 420
gccggtctgg caccggattg cgccattacc gttcatacgc aggcagatac cagctctccg 480
gacccgctgt tcaacccgct gaaaaccggc gtctgtcagc tggatgtcgc gcaagtgacg 540
gacgccattc tggaacgtgc aggcggttcc atcgctgatt ttaccggtca ctaccagacg 600
gcattccgtg aactggaacg cgttctgaac tttccgcagt caaatctggc gctgaaacgc 660
gaaaagcagg atgaaagtgc gtccctgacc caagccctgc cgagtgaact gaaagtctcc 720
gccgacaatg tgtcactgac cggcgcatgg tcactggctt cgatgctgac ggaaattttt 780
ctgctgcagc aagcacaggg tatgccggaa ccgggttggg gtcgtatcac cgattcgcat 840
cagtggaaca cgctgctgag cctgcacaat gcgcagttcg acctgctgca acgtaccccg 900
gaagtggcac gttcgcgcgc cacgccgctg ctggatctga ttaaaaccgc tctgacgccg 960
catccgccgc agaagcaagc gtatggcgtg accctgccga cgagcgttct gtttatcgcg 1020
ggtcacgaca ccaacctggc aaatctgggc ggtgctctgg aactgcagtg gaccctgccg 1080
ggtcaaccgg ataacacgcc gccgggcggt gaactggttt tcgaacgttg gcgtcgcctg 1140
agcgacaatt ctcagtggat ccaagttagc ctggtctttc agaccctgca gcaaatgcgc 1200
gataaaaccc cgctgttcct gaacacgccg ccgggcgaag tgaagctgac cctggcgggt 1260
tgcgaagaac gtaacgccca gggcatgtgt tctctggcag gttttaccca gattgttaat 1320
gaagcacgca tcccggcttg tagtctgtgt ctggacctga aaacgcaagt gcaaaccccg 1380
caaggcatga aggaaatctc aaacatccaa gtcggtgacc tggtgctgtc gaataccggc 1440
tataacgaag tgctgaatgt ttttccgaag agcaaaaaga aatcttacaa gatcacgctg 1500
gaagatggca aggaaattat ttgcagcgaa gaacatctgt tcccgaccca gacgggcgaa 1560
atgaatatct ccggcggtct gaaagaaggc atgtgtctgt acgtcaagga ataa 1614
<210> 92
<211> 537
<212> PRT
<213> Artificial Sequence
<400> 92
Met Met Leu Lys Lys Ile Leu Lys Ile Glu Glu Leu Asp Glu Arg Glu
1 5 10 15
Leu Ile Asp Ile Glu Val Ser Gly Asn His Leu Phe Tyr Ala Asn Asp
20 25 30
Ile Leu Thr His Asn Ser Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu
35 40 45
Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe
50 55 60
Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro
65 70 75 80
Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr
85 90 95
Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro
100 105 110
Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val
115 120 125
Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala
130 135 140
Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro
145 150 155 160
Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val
165 170 175
Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala
180 185 190
Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val
195 200 205
Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp
210 215 220
Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser
225 230 235 240
Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu
245 250 255
Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly
260 265 270
Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu
275 280 285
His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg
290 295 300
Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro
305 310 315 320
His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val
325 330 335
Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala
340 345 350
Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro
355 360 365
Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser
370 375 380
Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg
385 390 395 400
Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu
405 410 415
Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu
420 425 430
Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser
435 440 445
Leu Cys Leu Asp Leu Lys Thr Gln Val Gln Thr Pro Gln Gly Met Lys
450 455 460
Glu Ile Ser Asn Ile Gln Val Gly Asp Leu Val Leu Ser Asn Thr Gly
465 470 475 480
Tyr Asn Glu Val Leu Asn Val Phe Pro Lys Ser Lys Lys Lys Ser Tyr
485 490 495
Lys Ile Thr Leu Glu Asp Gly Lys Glu Ile Ile Cys Ser Glu Glu His
500 505 510
Leu Phe Pro Thr Gln Thr Gly Glu Met Asn Ile Ser Gly Gly Leu Lys
515 520 525
Glu Gly Met Cys Leu Tyr Val Lys Glu
530 535
<210> 93
<211> 536
<212> PRT
<213> Artificial Sequence
<400> 93
Met Met Leu Lys Lys Ile Leu Lys Ile Glu Glu Leu Asp Glu Arg Glu
1 5 10 15
Leu Ile Asp Ile Glu Val Ser Gly Asn His Leu Phe Tyr Ala Asn Asp
20 25 30
Ile Leu Thr His Asn Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe
35 40 45
Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro
50 55 60
Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr
65 70 75 80
Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro
85 90 95
Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val
100 105 110
Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala
115 120 125
Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro
130 135 140
Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val
145 150 155 160
Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala
165 170 175
Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val
180 185 190
Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp
195 200 205
Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser
210 215 220
Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu
225 230 235 240
Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly
245 250 255
Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu
260 265 270
His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg
275 280 285
Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro
290 295 300
His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val
305 310 315 320
Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala
325 330 335
Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro
340 345 350
Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser
355 360 365
Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg
370 375 380
Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu
385 390 395 400
Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu
405 410 415
Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser
420 425 430
Leu Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val Val Ile Val
435 440 445
Cys Leu Asp Leu Lys Thr Gln Val Gln Thr Pro Gln Gly Met Lys Glu
450 455 460
Ile Ser Asn Ile Gln Val Gly Asp Leu Val Leu Ser Asn Thr Gly Tyr
465 470 475 480
Asn Glu Val Leu Asn Val Phe Pro Lys Ser Lys Lys Lys Ser Tyr Lys
485 490 495
Ile Thr Leu Glu Asp Gly Lys Glu Ile Ile Cys Ser Glu Glu His Leu
500 505 510
Phe Pro Thr Gln Thr Gly Glu Met Asn Ile Ser Gly Gly Leu Lys Glu
515 520 525
Gly Met Cys Leu Tyr Val Lys Glu
530 535
<210> 94
<211> 1794
<212> DNA
<213> Artificial Sequence
<400> 94
atggttaagg tgattggaag acgttctctt ggtgttcaaa ggatcttcga tatcggattg 60
ccacaagacc acaactttct tctcgctaat ggtgccatcg ctgccaatag cgggggtggc 120
agtggaggcg gttcgacccc gcagtccgca tttgccgccc aatcggaacc ggaactgaaa 180
ctggaaagtg tggttattgt gtctcgtcat ggcgttcgcg ctccgaccaa atttacgcag 240
ctgatgcaag atgtcacccc ggacgccttc tatacgtggc cggtgaagct gggtgaactg 300
accccgcgtg gcggtgaact gatcgcctat ctgggtcact actggcgtca gcgcctggtg 360
gcagatggtc tgctgccgaa aaagggctgc ccgcagagcg gtcaagttgc aattatcgct 420
gatgtcgacg aacgtacccg caaaacgggt gaagcatttg cggccggtct ggcaccggat 480
tgcgccatta ccgttcatac gcaggcagat accagctctc cggacccgct gttcaacccg 540
ctgaaaaccg gcgtctgtca gctggatgtc gcgcaagtga cggacgccat tctggaacgt 600
gcaggcggtt ccatcgctga ttttaccggt cactaccaga cggcattccg tgaactggaa 660
cgcgttctga actttccgca gtcaaatctg gcgctgaaac gcgaaaagca ggatgaaagt 720
gcgtccctga cccaagccct gccgagtgaa ctgaaagtct ccgccgacaa tgtgtcactg 780
accggcgcat ggtcactggc ttcgatgctg acggaaattt ttctgctgca gcaagcacag 840
ggtatgccgg aaccgggttg gggtcgtatc accgattcgc atcagtggaa cacgctgctg 900
agcctgcaca atgcgcagtt cgacctgctg caacgtaccc cggaagtggc acgttcgcgc 960
gccacgccgc tgctggatct gattaaaacc gctctgacgc cgcatccgcc gcagaagcaa 1020
gcgtatggcg tgaccctgcc gacgagcgtt ctgtttatcg cgggtcacga caccaacctg 1080
gcaaatctgg gcggtgctct ggaactgcag tggaccctgc cgggtcaacc ggataacacg 1140
ccgccgggcg gtgaactggt tttcgaacgt tggcgtcgcc tgagcgacaa ttctcagtgg 1200
atccaagtta gcctggtctt tcagaccctg cagcaaatgc gcgataaaac cccgctgttc 1260
ctgaacacgc cgccgggcga agtgaagctg accctggcgg gttgcgaaga acgtaacgcc 1320
cagggcatgt gttctctggc aggttttacc cagattgtta atgaagcacg catcccggct 1380
tgtagtctgg gtggcgggag cggtggaggg agtgggggcg gttgcctttc tttcggaact 1440
gagatcctta ccgttgagta cggaccactt cctattggta agatcgtttc tgaggaaatt 1500
aactgctcag tgtactctgt tgatccagaa ggaagagttt acactcaggc tatcgcacaa 1560
tggcacgata ggggtgaaca agaggttctg gagtacgagc ttgaagatgg atccgttatt 1620
cgtgctacct ctgaccatag attcttgact acagattatc agcttctcgc tatcgaggaa 1680
atctttgcta ggcaacttga tctccttact ttggagaaca tcaagcagac agaagaggct 1740
cttgacaacc acagacttcc attccctttg ctcgatgctg gaaccatcaa gtaa 1794
<210> 95
<211> 597
<212> PRT
<213> Artificial Sequence
<400> 95
Met Val Lys Val Ile Gly Arg Arg Ser Leu Gly Val Gln Arg Ile Phe
1 5 10 15
Asp Ile Gly Leu Pro Gln Asp His Asn Phe Leu Leu Ala Asn Gly Ala
20 25 30
Ile Ala Ala Asn Ser Gly Gly Gly Ser Gly Gly Gly Ser Thr Pro Gln
35 40 45
Ser Ala Phe Ala Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val
50 55 60
Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe Thr Gln
65 70 75 80
Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro Val Lys
85 90 95
Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu Gly
100 105 110
His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys Lys
115 120 125
Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp Glu
130 135 140
Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro Asp
145 150 155 160
Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro Asp Pro
165 170 175
Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val Ala Gln
180 185 190
Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp Phe
195 200 205
Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu Asn
210 215 220
Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp Glu Ser
225 230 235 240
Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala Asp
245 250 255
Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu Thr Glu
260 265 270
Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp Gly
275 280 285
Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu His Asn
290 295 300
Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser Arg
305 310 315 320
Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His Pro
325 330 335
Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu Phe
340 345 350
Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu Glu
355 360 365
Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly
370 375 380
Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln Trp
385 390 395 400
Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp Lys
405 410 415
Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr Leu
420 425 430
Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala Gly
435 440 445
Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu Gly
450 455 460
Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Cys Leu Ser Phe Gly Thr
465 470 475 480
Glu Ile Leu Thr Val Glu Tyr Gly Pro Leu Pro Ile Gly Lys Ile Val
485 490 495
Ser Glu Glu Ile Asn Cys Ser Val Tyr Ser Val Asp Pro Glu Gly Arg
500 505 510
Val Tyr Thr Gln Ala Ile Ala Gln Trp His Asp Arg Gly Glu Gln Glu
515 520 525
Val Leu Glu Tyr Glu Leu Glu Asp Gly Ser Val Ile Arg Ala Thr Ser
530 535 540
Asp His Arg Phe Leu Thr Thr Asp Tyr Gln Leu Leu Ala Ile Glu Glu
545 550 555 560
Ile Phe Ala Arg Gln Leu Asp Leu Leu Thr Leu Glu Asn Ile Lys Gln
565 570 575
Thr Glu Glu Ala Leu Asp Asn His Arg Leu Pro Phe Pro Leu Leu Asp
580 585 590
Ala Gly Thr Ile Lys
595
<210> 96
<211> 1809
<212> DNA
<213> Artificial Sequence
<400> 96
atggttaagg tgattggaag acgttctctt ggtgttcaaa ggatcttcga tatcggattg 60
ccacaagacc acaactttct tctcgctaat ggtgccatcg ctgccaatag cggtggctcg 120
tcagggagta cgacaaccac gcgtatcacc ccgcaatctg cgttcgctgc ccaatcggaa 180
ccggaactga aactggaaag tgtggttatt gtgtctcgtc atggcgttcg cgctccgacc 240
aaatttacgc agctgatgca agatgtcacc ccggacgcct tctatacgtg gccggtgaag 300
ctgggtgaac tgaccccgcg tggcggtgaa ctgatcgcct atctgggtca ctactggcgt 360
cagcgcctgg tggcagatgg tctgctgccg aaaaagggct gcccgcagag cggtcaagtt 420
gcaattatcg ctgatgtcga cgaacgtacc cgcaaaacgg gtgaagcatt tgcggccggt 480
ctggcaccgg attgcgccat taccgttcat acgcaggcag ataccagctc tccggacccg 540
ctgttcaacc cgctgaaaac cggcgtctgt cagctggatg tcgcgcaagt gacggacgcc 600
attctggaac gtgcaggcgg ttccatcgct gattttaccg gtcactacca gacggcattc 660
cgtgaactgg aacgcgttct gaactttccg cagtcaaatc tggcgctgaa acgcgaaaag 720
caggatgaaa gtgcgtccct gacccaagcc ctgccgagtg aactgaaagt ctccgccgac 780
aatgtgtcac tgaccggcgc atggtcactg gcttcgatgc tgacggaaat ttttctgctg 840
cagcaagcac agggtatgcc ggaaccgggt tggggtcgta tcaccgattc gcatcagtgg 900
aacacgctgc tgagcctgca caatgcgcag ttcgacctgc tgcaacgtac cccggaagtg 960
gcacgttcgc gcgccacgcc gctgctggat ctgattaaaa ccgctctgac gccgcatccg 1020
ccgcagaagc aagcgtatgg cgtgaccctg ccgacgagcg ttctgtttat cgcgggtcac 1080
gacaccaacc tggcaaatct gggcggtgct ctggaactgc agtggaccct gccgggtcaa 1140
ccggataaca cgccgccggg cggtgaactg gttttcgaac gttggcgtcg cctgagcgac 1200
aattctcagt ggatccaagt tagcctggtc tttcagaccc tgcagcaaat gcgcgataaa 1260
accccgctgt tcctgaacac gccgccgggc gaagtgaagc tgaccctggc gggttgcgaa 1320
gaacgtaacg cccagggcat gtgttctctg gcaggtttta cccagattgt taatgaagca 1380
cgcatcccgg cttgtagtct gcaaaacacg tttagccagg ggagtagctc gggatcctgc 1440
ctttctttcg gaactgagat ccttaccgtt gagtacggac cacttcctat tggtaagatc 1500
gtttctgagg aaattaactg ctcagtgtac tctgttgatc cagaaggaag agtttacact 1560
caggctatcg cacaatggca cgataggggt gaacaagagg ttctggagta cgagcttgaa 1620
gatggatccg ttattcgtgc tacctctgac catagattct tgactacaga ttatcagctt 1680
ctcgctatcg aggaaatctt tgctaggcaa cttgatctcc ttactttgga gaacatcaag 1740
cagacagaag aggctcttga caaccacaga cttccattcc ctttgctcga tgctggaacc 1800
atcaagtaa 1809
<210> 97
<211> 602
<212> PRT
<213> Artificial Sequence
<400> 97
Met Val Lys Val Ile Gly Arg Arg Ser Leu Gly Val Gln Arg Ile Phe
1 5 10 15
Asp Ile Gly Leu Pro Gln Asp His Asn Phe Leu Leu Ala Asn Gly Ala
20 25 30
Ile Ala Ala Asn Ser Gly Gly Ser Ser Gly Ser Thr Thr Thr Thr Arg
35 40 45
Ile Thr Pro Gln Ser Ala Phe Ala Ala Gln Ser Glu Pro Glu Leu Lys
50 55 60
Leu Glu Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr
65 70 75 80
Lys Phe Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr
85 90 95
Trp Pro Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile
100 105 110
Ala Tyr Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu
115 120 125
Leu Pro Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala
130 135 140
Asp Val Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly
145 150 155 160
Leu Ala Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser
165 170 175
Ser Pro Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu
180 185 190
Asp Val Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser
195 200 205
Ile Ala Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu
210 215 220
Arg Val Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys
225 230 235 240
Gln Asp Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys
245 250 255
Val Ser Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser
260 265 270
Met Leu Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu
275 280 285
Pro Gly Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu
290 295 300
Ser Leu His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val
305 310 315 320
Ala Arg Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu
325 330 335
Thr Pro His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr
340 345 350
Ser Val Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly
355 360 365
Gly Ala Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr
370 375 380
Pro Pro Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp
385 390 395 400
Asn Ser Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln
405 410 415
Met Arg Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val
420 425 430
Lys Leu Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys
435 440 445
Ser Leu Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala
450 455 460
Cys Ser Leu Gln Asn Thr Phe Ser Gln Gly Ser Ser Ser Gly Ser Cys
465 470 475 480
Leu Ser Phe Gly Thr Glu Ile Leu Thr Val Glu Tyr Gly Pro Leu Pro
485 490 495
Ile Gly Lys Ile Val Ser Glu Glu Ile Asn Cys Ser Val Tyr Ser Val
500 505 510
Asp Pro Glu Gly Arg Val Tyr Thr Gln Ala Ile Ala Gln Trp His Asp
515 520 525
Arg Gly Glu Gln Glu Val Leu Glu Tyr Glu Leu Glu Asp Gly Ser Val
530 535 540
Ile Arg Ala Thr Ser Asp His Arg Phe Leu Thr Thr Asp Tyr Gln Leu
545 550 555 560
Leu Ala Ile Glu Glu Ile Phe Ala Arg Gln Leu Asp Leu Leu Thr Leu
565 570 575
Glu Asn Ile Lys Gln Thr Glu Glu Ala Leu Asp Asn His Arg Leu Pro
580 585 590
Phe Pro Leu Leu Asp Ala Gly Thr Ile Lys
595 600
<210> 98
<211> 1833
<212> DNA
<213> Artificial Sequence
<400> 98
atggttaagg tgattggaag acgttctctt ggtgttcaaa ggatcttcga tatcggattg 60
ccacaagacc acaactttct tctcgctaat ggtgccatcg ctgccaatag cgcctttgca 120
gcccaatcgg aaccggaact gaaactggaa agtgtggtta ttgtgtctcg tcatggcgtt 180
cgcgctccga ccaaatttac gcagctgatg caagatgtca ccccggacgc cttctatacg 240
tggccggtga agctgggtga actgaccccg cgtggcggtg aactgatcgc ctatctgggt 300
cactactggc gtcagcgcct ggtggcagat ggtctgctgc cgaaaaaggg ctgcccgcag 360
agcggtcaag ttgcaattat cgctgatgtc gacgaacgta cccgcaaaac gggtgaagca 420
tttgcggccg gtctggcacc ggattgcgcc attaccgttc atacgcaggc agataccagc 480
tctccggacc cgctgttcaa cccgctgaaa accggcgtct gtcagctgga tgtcgcgcaa 540
gtgacggacg ccattctgga acgtgcaggc ggttccatcg ctgattttac cggtcactac 600
cagacggcat tccgtgaact ggaacgcgtt ctgaactttc cgcagtcaaa tctggcgctg 660
aaacgcgaaa agcaggatga aagtgcgtcc ctgacccaag ccctgccgag tgaactgaaa 720
gtctccgccg acaatgtgtc actgaccggc gcatggtcac tggcttcgat gctgacggaa 780
atttttctgc tgcagcaagc acagggtatg ccggaaccgg gttggggtcg tatcaccgat 840
tcgcatcagt ggaacacgct gctgagcctg cacaatgcgc agttcgacct gctgcaacgt 900
accccggaag tggcacgttc gcgcgccacg ccgctgctgg atctgattaa aaccgctctg 960
acgccgcatc cgccgcagaa gcaagcgtat ggcgtgaccc tgccgacgag cgttctgttt 1020
atcgcgggtc acgacaccaa cctggcaaat ctgggcggtg ctctggaact gcagtggacc 1080
ctgccgggtc aaccggataa cacgccgccg ggcggtgaac tggttttcga acgttggcgt 1140
cgcctgagcg acaattctca gtggatccaa gttagcctgg tctttcagac cctgcagcaa 1200
atgcgcgata aaaccccgct gttcctgaac acgccgccgg gcgaagtgaa gctgaccctg 1260
gcgggttgcg aagaacgtaa cgcccagggc atgtgttctc tggcaggttt tacccagatt 1320
gttaatgaag cacgcatccc ggcttgtagt ctgggtgcag ctccagcggc cgcaccggct 1380
aaacaggaag cggcagctcc ggctcctgca gcgaaggcgg aagcaccggc cgcagctcct 1440
gcggcaaaag cgaccccgca gtgcctttct ttcggaactg agatccttac cgttgagtac 1500
ggaccacttc ctattggtaa gatcgtttct gaggaaatta actgctcagt gtactctgtt 1560
gatccagaag gaagagttta cactcaggct atcgcacaat ggcacgatag gggtgaacaa 1620
gaggttctgg agtacgagct tgaagatgga tccgttattc gtgctacctc tgaccataga 1680
ttcttgacta cagattatca gcttctcgct atcgaggaaa tctttgctag gcaacttgat 1740
ctccttactt tggagaacat caagcagaca gaagaggctc ttgacaacca cagacttcca 1800
ttccctttgc tcgatgctgg aaccatcaag taa 1833
<210> 99
<211> 610
<212> PRT
<213> Artificial Sequence
<400> 99
Met Val Lys Val Ile Gly Arg Arg Ser Leu Gly Val Gln Arg Ile Phe
1 5 10 15
Asp Ile Gly Leu Pro Gln Asp His Asn Phe Leu Leu Ala Asn Gly Ala
20 25 30
Ile Ala Ala Asn Ser Ala Phe Ala Ala Gln Ser Glu Pro Glu Leu Lys
35 40 45
Leu Glu Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr
50 55 60
Lys Phe Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr
65 70 75 80
Trp Pro Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile
85 90 95
Ala Tyr Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu
100 105 110
Leu Pro Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala
115 120 125
Asp Val Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly
130 135 140
Leu Ala Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser
145 150 155 160
Ser Pro Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu
165 170 175
Asp Val Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser
180 185 190
Ile Ala Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu
195 200 205
Arg Val Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys
210 215 220
Gln Asp Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys
225 230 235 240
Val Ser Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser
245 250 255
Met Leu Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu
260 265 270
Pro Gly Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu
275 280 285
Ser Leu His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val
290 295 300
Ala Arg Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu
305 310 315 320
Thr Pro His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr
325 330 335
Ser Val Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly
340 345 350
Gly Ala Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr
355 360 365
Pro Pro Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp
370 375 380
Asn Ser Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln
385 390 395 400
Met Arg Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val
405 410 415
Lys Leu Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys
420 425 430
Ser Leu Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala
435 440 445
Cys Ser Leu Gly Ala Ala Pro Ala Ala Ala Pro Ala Lys Gln Glu Ala
450 455 460
Ala Ala Pro Ala Pro Ala Ala Lys Ala Glu Ala Pro Ala Ala Ala Pro
465 470 475 480
Ala Ala Lys Ala Thr Pro Gln Cys Leu Ser Phe Gly Thr Glu Ile Leu
485 490 495
Thr Val Glu Tyr Gly Pro Leu Pro Ile Gly Lys Ile Val Ser Glu Glu
500 505 510
Ile Asn Cys Ser Val Tyr Ser Val Asp Pro Glu Gly Arg Val Tyr Thr
515 520 525
Gln Ala Ile Ala Gln Trp His Asp Arg Gly Glu Gln Glu Val Leu Glu
530 535 540
Tyr Glu Leu Glu Asp Gly Ser Val Ile Arg Ala Thr Ser Asp His Arg
545 550 555 560
Phe Leu Thr Thr Asp Tyr Gln Leu Leu Ala Ile Glu Glu Ile Phe Ala
565 570 575
Arg Gln Leu Asp Leu Leu Thr Leu Glu Asn Ile Lys Gln Thr Glu Glu
580 585 590
Ala Leu Asp Asn His Arg Leu Pro Phe Pro Leu Leu Asp Ala Gly Thr
595 600 605
Ile Lys
610
<210> 100
<211> 1860
<212> DNA
<213> Artificial Sequence
<400> 100
atggttaagg tgattggaag acgttctctt ggtgttcaaa ggatcttcga tatcggattg 60
ccacaagacc acaactttct tctcgctaat ggtgccatcg ctgccaatag cgcagccgaa 120
gccgctgcga aggaggcagc tgcgaaagaa gcggctgcaa aagaagcggc agctaaggct 180
ttgaataccc cgcaatcggc tttcgctgcc caatcggaac cggaactgaa actggaaagt 240
gtggttattg tgtctcgtca tggcgttcgc gctccgacca aatttacgca gctgatgcaa 300
gatgtcaccc cggacgcctt ctatacgtgg ccggtgaagc tgggtgaact gaccccgcgt 360
ggcggtgaac tgatcgccta tctgggtcac tactggcgtc agcgcctggt ggcagatggt 420
ctgctgccga aaaagggctg cccgcagagc ggtcaagttg caattatcgc tgatgtcgac 480
gaacgtaccc gcaaaacggg tgaagcattt gcggccggtc tggcaccgga ttgcgccatt 540
accgttcata cgcaggcaga taccagctct ccggacccgc tgttcaaccc gctgaaaacc 600
ggcgtctgtc agctggatgt cgcgcaagtg acggacgcca ttctggaacg tgcaggcggt 660
tccatcgctg attttaccgg tcactaccag acggcattcc gtgaactgga acgcgttctg 720
aactttccgc agtcaaatct ggcgctgaaa cgcgaaaagc aggatgaaag tgcgtccctg 780
acccaagccc tgccgagtga actgaaagtc tccgccgaca atgtgtcact gaccggcgca 840
tggtcactgg cttcgatgct gacggaaatt tttctgctgc agcaagcaca gggtatgccg 900
gaaccgggtt ggggtcgtat caccgattcg catcagtgga acacgctgct gagcctgcac 960
aatgcgcagt tcgacctgct gcaacgtacc ccggaagtgg cacgttcgcg cgccacgccg 1020
ctgctggatc tgattaaaac cgctctgacg ccgcatccgc cgcagaagca agcgtatggc 1080
gtgaccctgc cgacgagcgt tctgtttatc gcgggtcacg acaccaacct ggcaaatctg 1140
ggcggtgctc tggaactgca gtggaccctg ccgggtcaac cggataacac gccgccgggc 1200
ggtgaactgg ttttcgaacg ttggcgtcgc ctgagcgaca attctcagtg gatccaagtt 1260
agcctggtct ttcagaccct gcagcaaatg cgcgataaaa ccccgctgtt cctgaacacg 1320
ccgccgggcg aagtgaagct gaccctggcg ggttgcgaag aacgtaacgc ccagggcatg 1380
tgttctctgg caggttttac ccagattgtt aatgaagcac gcatcccggc ttgtagtctg 1440
gggggcgcag aagcagctgc caaagaggcg gccgcaaagg tcaatctgtg cctttctttc 1500
ggaactgaga tccttaccgt tgagtacgga ccacttccta ttggtaagat cgtttctgag 1560
gaaattaact gctcagtgta ctctgttgat ccagaaggaa gagtttacac tcaggctatc 1620
gcacaatggc acgatagggg tgaacaagag gttctggagt acgagcttga agatggatcc 1680
gttattcgtg ctacctctga ccatagattc ttgactacag attatcagct tctcgctatc 1740
gaggaaatct ttgctaggca acttgatctc cttactttgg agaacatcaa gcagacagaa 1800
gaggctcttg acaaccacag acttccattc cctttgctcg atgctggaac catcaagtaa 1860
<210> 101
<211> 619
<212> PRT
<213> Artificial Sequence
<400> 101
Met Val Lys Val Ile Gly Arg Arg Ser Leu Gly Val Gln Arg Ile Phe
1 5 10 15
Asp Ile Gly Leu Pro Gln Asp His Asn Phe Leu Leu Ala Asn Gly Ala
20 25 30
Ile Ala Ala Asn Ser Ala Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala
35 40 45
Lys Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Ala Leu Asn Thr Pro
50 55 60
Gln Ser Ala Phe Ala Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser
65 70 75 80
Val Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe Thr
85 90 95
Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro Val
100 105 110
Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu
115 120 125
Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys
130 135 140
Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp
145 150 155 160
Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro
165 170 175
Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro Asp
180 185 190
Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val Ala
195 200 205
Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp
210 215 220
Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu
225 230 235 240
Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp Glu
245 250 255
Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala
260 265 270
Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu Thr
275 280 285
Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp
290 295 300
Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu His
305 310 315 320
Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser
325 330 335
Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His
340 345 350
Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu
355 360 365
Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu
370 375 380
Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly
385 390 395 400
Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln
405 410 415
Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp
420 425 430
Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr
435 440 445
Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala
450 455 460
Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu
465 470 475 480
Gly Gly Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Val Asn Leu
485 490 495
Cys Leu Ser Phe Gly Thr Glu Ile Leu Thr Val Glu Tyr Gly Pro Leu
500 505 510
Pro Ile Gly Lys Ile Val Ser Glu Glu Ile Asn Cys Ser Val Tyr Ser
515 520 525
Val Asp Pro Glu Gly Arg Val Tyr Thr Gln Ala Ile Ala Gln Trp His
530 535 540
Asp Arg Gly Glu Gln Glu Val Leu Glu Tyr Glu Leu Glu Asp Gly Ser
545 550 555 560
Val Ile Arg Ala Thr Ser Asp His Arg Phe Leu Thr Thr Asp Tyr Gln
565 570 575
Leu Leu Ala Ile Glu Glu Ile Phe Ala Arg Gln Leu Asp Leu Leu Thr
580 585 590
Leu Glu Asn Ile Lys Gln Thr Glu Glu Ala Leu Asp Asn His Arg Leu
595 600 605
Pro Phe Pro Leu Leu Asp Ala Gly Thr Ile Lys
610 615
<210> 102
<211> 1503
<212> DNA
<213> Artificial Sequence
<400> 102
atgagggcca agcagctgga ggacaagatt gaggagctgc tgagcaagat ctaccacctg 60
gagaacgaga tagcccgcct gaagaagctg attggcgagc gcagcggggg tggcagtgga 120
ggcggttcga ccccgcagtc cgcatttgcc gcccaatcgg aaccggaact gaaactggaa 180
agtgtggtta ttgtgtctcg tcatggcgtt cgcgctccga ccaaatttac gcagctgatg 240
caagatgtca ccccggacgc cttctatacg tggccggtga agctgggtga actgaccccg 300
cgtggcggtg aactgatcgc ctatctgggt cactactggc gtcagcgcct ggtggcagat 360
ggtctgctgc cgaaaaaggg ctgcccgcag agcggtcaag ttgcaattat cgctgatgtc 420
gacgaacgta cccgcaaaac gggtgaagca tttgcggccg gtctggcacc ggattgcgcc 480
attaccgttc atacgcaggc agataccagc tctccggacc cgctgttcaa cccgctgaaa 540
accggcgtct gtcagctgga tgtcgcgcaa gtgacggacg ccattctgga acgtgcaggc 600
ggttccatcg ctgattttac cggtcactac cagacggcat tccgtgaact ggaacgcgtt 660
ctgaactttc cgcagtcaaa tctggcgctg aaacgcgaaa agcaggatga aagtgcgtcc 720
ctgacccaag ccctgccgag tgaactgaaa gtctccgccg acaatgtgtc actgaccggc 780
gcatggtcac tggcttcgat gctgacggaa atttttctgc tgcagcaagc acagggtatg 840
ccggaaccgg gttggggtcg tatcaccgat tcgcatcagt ggaacacgct gctgagcctg 900
cacaatgcgc agttcgacct gctgcaacgt accccggaag tggcacgttc gcgcgccacg 960
ccgctgctgg atctgattaa aaccgctctg acgccgcatc cgccgcagaa gcaagcgtat 1020
ggcgtgaccc tgccgacgag cgttctgttt atcgcgggtc acgacaccaa cctggcaaat 1080
ctgggcggtg ctctggaact gcagtggacc ctgccgggtc aaccggataa cacgccgccg 1140
ggcggtgaac tggttttcga acgttggcgt cgcctgagcg acaattctca gtggatccaa 1200
gttagcctgg tctttcagac cctgcagcaa atgcgcgata aaaccccgct gttcctgaac 1260
acgccgccgg gcgaagtgaa gctgaccctg gcgggttgcg aagaacgtaa cgcccagggc 1320
atgtgttctc tggcaggttt tacccagatt gttaatgaag cacgcatccc ggcttgtagt 1380
ctgggtggcg ggagcggtgg agggagtggg ggcggtcagc tggaggacaa gattgaggag 1440
ctgctgagca agatctacca cctggagaac gagatagcga ggctgaagaa gctgattggc 1500
taa 1503
<210> 103
<211> 500
<212> PRT
<213> Artificial Sequence
<400> 103
Met Arg Ala Lys Gln Leu Glu Asp Lys Ile Glu Glu Leu Leu Ser Lys
1 5 10 15
Ile Tyr His Leu Glu Asn Glu Ile Ala Arg Leu Lys Lys Leu Ile Gly
20 25 30
Glu Arg Ser Gly Gly Gly Ser Gly Gly Gly Ser Thr Pro Gln Ser Ala
35 40 45
Phe Ala Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val Val Ile
50 55 60
Val Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe Thr Gln Leu Met
65 70 75 80
Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro Val Lys Leu Gly
85 90 95
Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu Gly His Tyr
100 105 110
Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys Lys Gly Cys
115 120 125
Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp Glu Arg Thr
130 135 140
Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro Asp Cys Ala
145 150 155 160
Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro Asp Pro Leu Phe
165 170 175
Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val Ala Gln Val Thr
180 185 190
Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp Phe Thr Gly
195 200 205
His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu Asn Phe Pro
210 215 220
Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp Glu Ser Ala Ser
225 230 235 240
Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala Asp Asn Val
245 250 255
Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu Thr Glu Ile Phe
260 265 270
Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp Gly Arg Ile
275 280 285
Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu His Asn Ala Gln
290 295 300
Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser Arg Ala Thr
305 310 315 320
Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His Pro Pro Gln
325 330 335
Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu Phe Ile Ala
340 345 350
Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu Glu Leu Gln
355 360 365
Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly Glu Leu
370 375 380
Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln Trp Ile Gln
385 390 395 400
Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp Lys Thr Pro
405 410 415
Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr Leu Ala Gly
420 425 430
Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala Gly Phe Thr
435 440 445
Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu Gly Gly Gly
450 455 460
Ser Gly Gly Gly Ser Gly Gly Gly Gln Leu Glu Asp Lys Ile Glu Glu
465 470 475 480
Leu Leu Ser Lys Ile Tyr His Leu Glu Asn Glu Ile Ala Arg Leu Lys
485 490 495
Lys Leu Ile Gly
500
<210> 104
<211> 1518
<212> DNA
<213> Artificial Sequence
<400> 104
atgagggcca agcagctgga ggacaagatt gaggagctgc tgagcaagat ctaccacctg 60
gagaacgaga tagcccgcct gaagaagctg attggcgagc gcagcggtgg ctcgtcaggg 120
agtacgacaa ccacgcgtat caccccgcaa tctgcgttcg ctgcccaatc ggaaccggaa 180
ctgaaactgg aaagtgtggt tattgtgtct cgtcatggcg ttcgcgctcc gaccaaattt 240
acgcagctga tgcaagatgt caccccggac gccttctata cgtggccggt gaagctgggt 300
gaactgaccc cgcgtggcgg tgaactgatc gcctatctgg gtcactactg gcgtcagcgc 360
ctggtggcag atggtctgct gccgaaaaag ggctgcccgc agagcggtca agttgcaatt 420
atcgctgatg tcgacgaacg tacccgcaaa acgggtgaag catttgcggc cggtctggca 480
ccggattgcg ccattaccgt tcatacgcag gcagatacca gctctccgga cccgctgttc 540
aacccgctga aaaccggcgt ctgtcagctg gatgtcgcgc aagtgacgga cgccattctg 600
gaacgtgcag gcggttccat cgctgatttt accggtcact accagacggc attccgtgaa 660
ctggaacgcg ttctgaactt tccgcagtca aatctggcgc tgaaacgcga aaagcaggat 720
gaaagtgcgt ccctgaccca agccctgccg agtgaactga aagtctccgc cgacaatgtg 780
tcactgaccg gcgcatggtc actggcttcg atgctgacgg aaatttttct gctgcagcaa 840
gcacagggta tgccggaacc gggttggggt cgtatcaccg attcgcatca gtggaacacg 900
ctgctgagcc tgcacaatgc gcagttcgac ctgctgcaac gtaccccgga agtggcacgt 960
tcgcgcgcca cgccgctgct ggatctgatt aaaaccgctc tgacgccgca tccgccgcag 1020
aagcaagcgt atggcgtgac cctgccgacg agcgttctgt ttatcgcggg tcacgacacc 1080
aacctggcaa atctgggcgg tgctctggaa ctgcagtgga ccctgccggg tcaaccggat 1140
aacacgccgc cgggcggtga actggttttc gaacgttggc gtcgcctgag cgacaattct 1200
cagtggatcc aagttagcct ggtctttcag accctgcagc aaatgcgcga taaaaccccg 1260
ctgttcctga acacgccgcc gggcgaagtg aagctgaccc tggcgggttg cgaagaacgt 1320
aacgcccagg gcatgtgttc tctggcaggt tttacccaga ttgttaatga agcacgcatc 1380
ccggcttgta gtctgcaaaa cacgtttagc caggggagta gctcgggatc ccagctggag 1440
gacaagattg aggagctgct gagcaagatc taccacctgg agaacgagat agcgaggctg 1500
aagaagctga ttggctaa 1518
<210> 105
<211> 505
<212> PRT
<213> Artificial Sequence
<400> 105
Met Arg Ala Lys Gln Leu Glu Asp Lys Ile Glu Glu Leu Leu Ser Lys
1 5 10 15
Ile Tyr His Leu Glu Asn Glu Ile Ala Arg Leu Lys Lys Leu Ile Gly
20 25 30
Glu Arg Ser Gly Gly Ser Ser Gly Ser Thr Thr Thr Thr Arg Ile Thr
35 40 45
Pro Gln Ser Ala Phe Ala Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu
50 55 60
Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe
65 70 75 80
Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro
85 90 95
Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr
100 105 110
Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro
115 120 125
Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val
130 135 140
Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala
145 150 155 160
Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro
165 170 175
Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val
180 185 190
Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala
195 200 205
Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val
210 215 220
Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp
225 230 235 240
Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser
245 250 255
Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu
260 265 270
Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly
275 280 285
Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu
290 295 300
His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg
305 310 315 320
Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro
325 330 335
His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val
340 345 350
Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala
355 360 365
Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro
370 375 380
Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser
385 390 395 400
Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg
405 410 415
Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu
420 425 430
Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu
435 440 445
Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser
450 455 460
Leu Gln Asn Thr Phe Ser Gln Gly Ser Ser Ser Gly Ser Gln Leu Glu
465 470 475 480
Asp Lys Ile Glu Glu Leu Leu Ser Lys Ile Tyr His Leu Glu Asn Glu
485 490 495
Ile Ala Arg Leu Lys Lys Leu Ile Gly
500 505
<210> 106
<211> 1542
<212> DNA
<213> Artificial Sequence
<400> 106
atgagggcca agcagctgga ggacaagatt gaggagctgc tgagcaagat ctaccacctg 60
gagaacgaga tagcccgcct gaagaagctg attggcgagc gcagcgcctt tgcagcccaa 120
tcggaaccgg aactgaaact ggaaagtgtg gttattgtgt ctcgtcatgg cgttcgcgct 180
ccgaccaaat ttacgcagct gatgcaagat gtcaccccgg acgccttcta tacgtggccg 240
gtgaagctgg gtgaactgac cccgcgtggc ggtgaactga tcgcctatct gggtcactac 300
tggcgtcagc gcctggtggc agatggtctg ctgccgaaaa agggctgccc gcagagcggt 360
caagttgcaa ttatcgctga tgtcgacgaa cgtacccgca aaacgggtga agcatttgcg 420
gccggtctgg caccggattg cgccattacc gttcatacgc aggcagatac cagctctccg 480
gacccgctgt tcaacccgct gaaaaccggc gtctgtcagc tggatgtcgc gcaagtgacg 540
gacgccattc tggaacgtgc aggcggttcc atcgctgatt ttaccggtca ctaccagacg 600
gcattccgtg aactggaacg cgttctgaac tttccgcagt caaatctggc gctgaaacgc 660
gaaaagcagg atgaaagtgc gtccctgacc caagccctgc cgagtgaact gaaagtctcc 720
gccgacaatg tgtcactgac cggcgcatgg tcactggctt cgatgctgac ggaaattttt 780
ctgctgcagc aagcacaggg tatgccggaa ccgggttggg gtcgtatcac cgattcgcat 840
cagtggaaca cgctgctgag cctgcacaat gcgcagttcg acctgctgca acgtaccccg 900
gaagtggcac gttcgcgcgc cacgccgctg ctggatctga ttaaaaccgc tctgacgccg 960
catccgccgc agaagcaagc gtatggcgtg accctgccga cgagcgttct gtttatcgcg 1020
ggtcacgaca ccaacctggc aaatctgggc ggtgctctgg aactgcagtg gaccctgccg 1080
ggtcaaccgg ataacacgcc gccgggcggt gaactggttt tcgaacgttg gcgtcgcctg 1140
agcgacaatt ctcagtggat ccaagttagc ctggtctttc agaccctgca gcaaatgcgc 1200
gataaaaccc cgctgttcct gaacacgccg ccgggcgaag tgaagctgac cctggcgggt 1260
tgcgaagaac gtaacgccca gggcatgtgt tctctggcag gttttaccca gattgttaat 1320
gaagcacgca tcccggcttg tagtctgggt gcagctccag cggccgcacc ggctaaacag 1380
gaagcggcag ctccggctcc tgcagcgaag gcggaagcac cggccgcagc tcctgcggca 1440
aaagcgaccc cgcagcagct ggaggacaag attgaggagc tgctgagcaa gatctaccac 1500
ctggagaacg agatagcgag gctgaagaag ctgattggct aa 1542
<210> 107
<211> 513
<212> PRT
<213> Artificial Sequence
<400> 107
Met Arg Ala Lys Gln Leu Glu Asp Lys Ile Glu Glu Leu Leu Ser Lys
1 5 10 15
Ile Tyr His Leu Glu Asn Glu Ile Ala Arg Leu Lys Lys Leu Ile Gly
20 25 30
Glu Arg Ser Ala Phe Ala Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu
35 40 45
Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe
50 55 60
Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro
65 70 75 80
Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr
85 90 95
Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro
100 105 110
Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val
115 120 125
Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala
130 135 140
Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro
145 150 155 160
Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val
165 170 175
Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala
180 185 190
Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val
195 200 205
Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp
210 215 220
Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser
225 230 235 240
Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu
245 250 255
Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly
260 265 270
Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu
275 280 285
His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg
290 295 300
Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro
305 310 315 320
His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val
325 330 335
Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala
340 345 350
Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro
355 360 365
Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser
370 375 380
Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg
385 390 395 400
Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu
405 410 415
Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu
420 425 430
Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser
435 440 445
Leu Gly Ala Ala Pro Ala Ala Ala Pro Ala Lys Gln Glu Ala Ala Ala
450 455 460
Pro Ala Pro Ala Ala Lys Ala Glu Ala Pro Ala Ala Ala Pro Ala Ala
465 470 475 480
Lys Ala Thr Pro Gln Gln Leu Glu Asp Lys Ile Glu Glu Leu Leu Ser
485 490 495
Lys Ile Tyr His Leu Glu Asn Glu Ile Ala Arg Leu Lys Lys Leu Ile
500 505 510
Gly
<210> 108
<211> 1569
<212> DNA
<213> Artificial Sequence
<400> 108
atgagggcca agcagctgga ggacaagatt gaggagctgc tgagcaagat ctaccacctg 60
gagaacgaga tagcccgcct gaagaagctg attggcgagc gcagcgcagc cgaagccgct 120
gcgaaggagg cagctgcgaa agaagcggct gcaaaagaag cggcagctaa ggctttgaat 180
accccgcaat cggctttcgc tgcccaatcg gaaccggaac tgaaactgga aagtgtggtt 240
attgtgtctc gtcatggcgt tcgcgctccg accaaattta cgcagctgat gcaagatgtc 300
accccggacg ccttctatac gtggccggtg aagctgggtg aactgacccc gcgtggcggt 360
gaactgatcg cctatctggg tcactactgg cgtcagcgcc tggtggcaga tggtctgctg 420
ccgaaaaagg gctgcccgca gagcggtcaa gttgcaatta tcgctgatgt cgacgaacgt 480
acccgcaaaa cgggtgaagc atttgcggcc ggtctggcac cggattgcgc cattaccgtt 540
catacgcagg cagataccag ctctccggac ccgctgttca acccgctgaa aaccggcgtc 600
tgtcagctgg atgtcgcgca agtgacggac gccattctgg aacgtgcagg cggttccatc 660
gctgatttta ccggtcacta ccagacggca ttccgtgaac tggaacgcgt tctgaacttt 720
ccgcagtcaa atctggcgct gaaacgcgaa aagcaggatg aaagtgcgtc cctgacccaa 780
gccctgccga gtgaactgaa agtctccgcc gacaatgtgt cactgaccgg cgcatggtca 840
ctggcttcga tgctgacgga aatttttctg ctgcagcaag cacagggtat gccggaaccg 900
ggttggggtc gtatcaccga ttcgcatcag tggaacacgc tgctgagcct gcacaatgcg 960
cagttcgacc tgctgcaacg taccccggaa gtggcacgtt cgcgcgccac gccgctgctg 1020
gatctgatta aaaccgctct gacgccgcat ccgccgcaga agcaagcgta tggcgtgacc 1080
ctgccgacga gcgttctgtt tatcgcgggt cacgacacca acctggcaaa tctgggcggt 1140
gctctggaac tgcagtggac cctgccgggt caaccggata acacgccgcc gggcggtgaa 1200
ctggttttcg aacgttggcg tcgcctgagc gacaattctc agtggatcca agttagcctg 1260
gtctttcaga ccctgcagca aatgcgcgat aaaaccccgc tgttcctgaa cacgccgccg 1320
ggcgaagtga agctgaccct ggcgggttgc gaagaacgta acgcccaggg catgtgttct 1380
ctggcaggtt ttacccagat tgttaatgaa gcacgcatcc cggcttgtag tctggggggc 1440
gcagaagcag ctgccaaaga ggcggccgca aaggtcaatc tgcagctgga ggacaagatt 1500
gaggagctgc tgagcaagat ctaccacctg gagaacgaga tagcgaggct gaagaagctg 1560
attggctaa 1569
<210> 109
<211> 522
<212> PRT
<213> Artificial Sequence
<400> 109
Met Arg Ala Lys Gln Leu Glu Asp Lys Ile Glu Glu Leu Leu Ser Lys
1 5 10 15
Ile Tyr His Leu Glu Asn Glu Ile Ala Arg Leu Lys Lys Leu Ile Gly
20 25 30
Glu Arg Ser Ala Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Glu
35 40 45
Ala Ala Ala Lys Glu Ala Ala Ala Lys Ala Leu Asn Thr Pro Gln Ser
50 55 60
Ala Phe Ala Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val Val
65 70 75 80
Ile Val Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe Thr Gln Leu
85 90 95
Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro Val Lys Leu
100 105 110
Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu Gly His
115 120 125
Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys Lys Gly
130 135 140
Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp Glu Arg
145 150 155 160
Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro Asp Cys
165 170 175
Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro Asp Pro Leu
180 185 190
Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val Ala Gln Val
195 200 205
Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp Phe Thr
210 215 220
Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu Asn Phe
225 230 235 240
Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp Glu Ser Ala
245 250 255
Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala Asp Asn
260 265 270
Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu Thr Glu Ile
275 280 285
Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp Gly Arg
290 295 300
Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu His Asn Ala
305 310 315 320
Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser Arg Ala
325 330 335
Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His Pro Pro
340 345 350
Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu Phe Ile
355 360 365
Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu Glu Leu
370 375 380
Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly Glu
385 390 395 400
Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln Trp Ile
405 410 415
Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp Lys Thr
420 425 430
Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr Leu Ala
435 440 445
Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala Gly Phe
450 455 460
Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu Gly Gly
465 470 475 480
Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Val Asn Leu Gln Leu
485 490 495
Glu Asp Lys Ile Glu Glu Leu Leu Ser Lys Ile Tyr His Leu Glu Asn
500 505 510
Glu Ile Ala Arg Leu Lys Lys Leu Ile Gly
515 520
<210> 110
<211> 1503
<212> DNA
<213> Artificial Sequence
<400> 110
atgagggcca agcagctgga ggacaaggtc gaggagctgc tgagcaagaa ctaccacctg 60
gagaacgagg tcgcccgcct gaagaagctg gtgggcaccc gcagcggggg tggcagtgga 120
ggcggttcga ccccgcagtc cgcatttgcc gcccaatcgg aaccggaact gaaactggaa 180
agtgtggtta ttgtgtctcg tcatggcgtt cgcgctccga ccaaatttac gcagctgatg 240
caagatgtca ccccggacgc cttctatacg tggccggtga agctgggtga actgaccccg 300
cgtggcggtg aactgatcgc ctatctgggt cactactggc gtcagcgcct ggtggcagat 360
ggtctgctgc cgaaaaaggg ctgcccgcag agcggtcaag ttgcaattat cgctgatgtc 420
gacgaacgta cccgcaaaac gggtgaagca tttgcggccg gtctggcacc ggattgcgcc 480
attaccgttc atacgcaggc agataccagc tctccggacc cgctgttcaa cccgctgaaa 540
accggcgtct gtcagctgga tgtcgcgcaa gtgacggacg ccattctgga acgtgcaggc 600
ggttccatcg ctgattttac cggtcactac cagacggcat tccgtgaact ggaacgcgtt 660
ctgaactttc cgcagtcaaa tctggcgctg aaacgcgaaa agcaggatga aagtgcgtcc 720
ctgacccaag ccctgccgag tgaactgaaa gtctccgccg acaatgtgtc actgaccggc 780
gcatggtcac tggcttcgat gctgacggaa atttttctgc tgcagcaagc acagggtatg 840
ccggaaccgg gttggggtcg tatcaccgat tcgcatcagt ggaacacgct gctgagcctg 900
cacaatgcgc agttcgacct gctgcaacgt accccggaag tggcacgttc gcgcgccacg 960
ccgctgctgg atctgattaa aaccgctctg acgccgcatc cgccgcagaa gcaagcgtat 1020
ggcgtgaccc tgccgacgag cgttctgttt atcgcgggtc acgacaccaa cctggcaaat 1080
ctgggcggtg ctctggaact gcagtggacc ctgccgggtc aaccggataa cacgccgccg 1140
ggcggtgaac tggttttcga acgttggcgt cgcctgagcg acaattctca gtggatccaa 1200
gttagcctgg tctttcagac cctgcagcaa atgcgcgata aaaccccgct gttcctgaac 1260
acgccgccgg gcgaagtgaa gctgaccctg gcgggttgcg aagaacgtaa cgcccagggc 1320
atgtgttctc tggcaggttt tacccagatt gttaatgaag cacgcatccc ggcttgtagt 1380
ctgggtggcg ggagcggtgg agggagtggg ggcggtcaat tggaagataa agtggaagag 1440
ctcctgtcca aaaattatca tctggaaaat gaggtggccc gcttgaagaa actcgtggga 1500
taa 1503
<210> 111
<211> 500
<212> PRT
<213> Artificial Sequence
<400> 111
Met Arg Ala Lys Gln Leu Glu Asp Lys Val Glu Glu Leu Leu Ser Lys
1 5 10 15
Asn Tyr His Leu Glu Asn Glu Val Ala Arg Leu Lys Lys Leu Val Gly
20 25 30
Thr Arg Ser Gly Gly Gly Ser Gly Gly Gly Ser Thr Pro Gln Ser Ala
35 40 45
Phe Ala Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val Val Ile
50 55 60
Val Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe Thr Gln Leu Met
65 70 75 80
Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro Val Lys Leu Gly
85 90 95
Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu Gly His Tyr
100 105 110
Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys Lys Gly Cys
115 120 125
Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp Glu Arg Thr
130 135 140
Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro Asp Cys Ala
145 150 155 160
Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro Asp Pro Leu Phe
165 170 175
Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val Ala Gln Val Thr
180 185 190
Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp Phe Thr Gly
195 200 205
His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu Asn Phe Pro
210 215 220
Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp Glu Ser Ala Ser
225 230 235 240
Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala Asp Asn Val
245 250 255
Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu Thr Glu Ile Phe
260 265 270
Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp Gly Arg Ile
275 280 285
Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu His Asn Ala Gln
290 295 300
Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser Arg Ala Thr
305 310 315 320
Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His Pro Pro Gln
325 330 335
Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu Phe Ile Ala
340 345 350
Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu Glu Leu Gln
355 360 365
Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly Glu Leu
370 375 380
Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln Trp Ile Gln
385 390 395 400
Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp Lys Thr Pro
405 410 415
Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr Leu Ala Gly
420 425 430
Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala Gly Phe Thr
435 440 445
Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu Gly Gly Gly
450 455 460
Ser Gly Gly Gly Ser Gly Gly Gly Gln Leu Glu Asp Lys Val Glu Glu
465 470 475 480
Leu Leu Ser Lys Asn Tyr His Leu Glu Asn Glu Val Ala Arg Leu Lys
485 490 495
Lys Leu Val Gly
500
<210> 112
<211> 1518
<212> DNA
<213> Artificial Sequence
<400> 112
atgagggcca agcagctgga ggacaaggtc gaggagctgc tgagcaagaa ctaccacctg 60
gagaacgagg tcgcccgcct gaagaagctg gtgggcaccc gcagcggtgg ctcgtcaggg 120
agtacgacaa ccacgcgtat caccccgcaa tctgcgttcg ctgcccaatc ggaaccggaa 180
ctgaaactgg aaagtgtggt tattgtgtct cgtcatggcg ttcgcgctcc gaccaaattt 240
acgcagctga tgcaagatgt caccccggac gccttctata cgtggccggt gaagctgggt 300
gaactgaccc cgcgtggcgg tgaactgatc gcctatctgg gtcactactg gcgtcagcgc 360
ctggtggcag atggtctgct gccgaaaaag ggctgcccgc agagcggtca agttgcaatt 420
atcgctgatg tcgacgaacg tacccgcaaa acgggtgaag catttgcggc cggtctggca 480
ccggattgcg ccattaccgt tcatacgcag gcagatacca gctctccgga cccgctgttc 540
aacccgctga aaaccggcgt ctgtcagctg gatgtcgcgc aagtgacgga cgccattctg 600
gaacgtgcag gcggttccat cgctgatttt accggtcact accagacggc attccgtgaa 660
ctggaacgcg ttctgaactt tccgcagtca aatctggcgc tgaaacgcga aaagcaggat 720
gaaagtgcgt ccctgaccca agccctgccg agtgaactga aagtctccgc cgacaatgtg 780
tcactgaccg gcgcatggtc actggcttcg atgctgacgg aaatttttct gctgcagcaa 840
gcacagggta tgccggaacc gggttggggt cgtatcaccg attcgcatca gtggaacacg 900
ctgctgagcc tgcacaatgc gcagttcgac ctgctgcaac gtaccccgga agtggcacgt 960
tcgcgcgcca cgccgctgct ggatctgatt aaaaccgctc tgacgccgca tccgccgcag 1020
aagcaagcgt atggcgtgac cctgccgacg agcgttctgt ttatcgcggg tcacgacacc 1080
aacctggcaa atctgggcgg tgctctggaa ctgcagtgga ccctgccggg tcaaccggat 1140
aacacgccgc cgggcggtga actggttttc gaacgttggc gtcgcctgag cgacaattct 1200
cagtggatcc aagttagcct ggtctttcag accctgcagc aaatgcgcga taaaaccccg 1260
ctgttcctga acacgccgcc gggcgaagtg aagctgaccc tggcgggttg cgaagaacgt 1320
aacgcccagg gcatgtgttc tctggcaggt tttacccaga ttgttaatga agcacgcatc 1380
ccggcttgta gtctgcaaaa cacgtttagc caggggagta gctcgggatc ccaattggaa 1440
gataaagtgg aagagctcct gtccaaaaat tatcatctgg aaaatgaggt ggcccgcttg 1500
aagaaactcg tgggataa 1518
<210> 113
<211> 505
<212> PRT
<213> Artificial Sequence
<400> 113
Met Arg Ala Lys Gln Leu Glu Asp Lys Val Glu Glu Leu Leu Ser Lys
1 5 10 15
Asn Tyr His Leu Glu Asn Glu Val Ala Arg Leu Lys Lys Leu Val Gly
20 25 30
Thr Arg Ser Gly Gly Ser Ser Gly Ser Thr Thr Thr Thr Arg Ile Thr
35 40 45
Pro Gln Ser Ala Phe Ala Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu
50 55 60
Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe
65 70 75 80
Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro
85 90 95
Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr
100 105 110
Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro
115 120 125
Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val
130 135 140
Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala
145 150 155 160
Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro
165 170 175
Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val
180 185 190
Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala
195 200 205
Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val
210 215 220
Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp
225 230 235 240
Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser
245 250 255
Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu
260 265 270
Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly
275 280 285
Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu
290 295 300
His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg
305 310 315 320
Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro
325 330 335
His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val
340 345 350
Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala
355 360 365
Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro
370 375 380
Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser
385 390 395 400
Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg
405 410 415
Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu
420 425 430
Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu
435 440 445
Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser
450 455 460
Leu Gln Asn Thr Phe Ser Gln Gly Ser Ser Ser Gly Ser Gln Leu Glu
465 470 475 480
Asp Lys Val Glu Glu Leu Leu Ser Lys Asn Tyr His Leu Glu Asn Glu
485 490 495
Val Ala Arg Leu Lys Lys Leu Val Gly
500 505
<210> 114
<211> 1542
<212> DNA
<213> Artificial Sequence
<400> 114
atgagggcca agcagctgga ggacaaggtc gaggagctgc tgagcaagaa ctaccacctg 60
gagaacgagg tcgcccgcct gaagaagctg gtgggcaccc gcagcgcctt tgcagcccaa 120
tcggaaccgg aactgaaact ggaaagtgtg gttattgtgt ctcgtcatgg cgttcgcgct 180
ccgaccaaat ttacgcagct gatgcaagat gtcaccccgg acgccttcta tacgtggccg 240
gtgaagctgg gtgaactgac cccgcgtggc ggtgaactga tcgcctatct gggtcactac 300
tggcgtcagc gcctggtggc agatggtctg ctgccgaaaa agggctgccc gcagagcggt 360
caagttgcaa ttatcgctga tgtcgacgaa cgtacccgca aaacgggtga agcatttgcg 420
gccggtctgg caccggattg cgccattacc gttcatacgc aggcagatac cagctctccg 480
gacccgctgt tcaacccgct gaaaaccggc gtctgtcagc tggatgtcgc gcaagtgacg 540
gacgccattc tggaacgtgc aggcggttcc atcgctgatt ttaccggtca ctaccagacg 600
gcattccgtg aactggaacg cgttctgaac tttccgcagt caaatctggc gctgaaacgc 660
gaaaagcagg atgaaagtgc gtccctgacc caagccctgc cgagtgaact gaaagtctcc 720
gccgacaatg tgtcactgac cggcgcatgg tcactggctt cgatgctgac ggaaattttt 780
ctgctgcagc aagcacaggg tatgccggaa ccgggttggg gtcgtatcac cgattcgcat 840
cagtggaaca cgctgctgag cctgcacaat gcgcagttcg acctgctgca acgtaccccg 900
gaagtggcac gttcgcgcgc cacgccgctg ctggatctga ttaaaaccgc tctgacgccg 960
catccgccgc agaagcaagc gtatggcgtg accctgccga cgagcgttct gtttatcgcg 1020
ggtcacgaca ccaacctggc aaatctgggc ggtgctctgg aactgcagtg gaccctgccg 1080
ggtcaaccgg ataacacgcc gccgggcggt gaactggttt tcgaacgttg gcgtcgcctg 1140
agcgacaatt ctcagtggat ccaagttagc ctggtctttc agaccctgca gcaaatgcgc 1200
gataaaaccc cgctgttcct gaacacgccg ccgggcgaag tgaagctgac cctggcgggt 1260
tgcgaagaac gtaacgccca gggcatgtgt tctctggcag gttttaccca gattgttaat 1320
gaagcacgca tcccggcttg tagtctgggt gcagctccag cggccgcacc ggctaaacag 1380
gaagcggcag ctccggctcc tgcagcgaag gcggaagcac cggccgcagc tcctgcggca 1440
aaagcgaccc cgcagcaatt ggaagataaa gtggaagagc tcctgtccaa aaattatcat 1500
ctggaaaatg aggtggcccg cttgaagaaa ctcgtgggat aa 1542
<210> 115
<211> 513
<212> PRT
<213> Artificial Sequence
<400> 115
Met Arg Ala Lys Gln Leu Glu Asp Lys Val Glu Glu Leu Leu Ser Lys
1 5 10 15
Asn Tyr His Leu Glu Asn Glu Val Ala Arg Leu Lys Lys Leu Val Gly
20 25 30
Thr Arg Ser Ala Phe Ala Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu
35 40 45
Ser Val Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe
50 55 60
Thr Gln Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro
65 70 75 80
Val Lys Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr
85 90 95
Leu Gly His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro
100 105 110
Lys Lys Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val
115 120 125
Asp Glu Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala
130 135 140
Pro Asp Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro
145 150 155 160
Asp Pro Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val
165 170 175
Ala Gln Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala
180 185 190
Asp Phe Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val
195 200 205
Leu Asn Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp
210 215 220
Glu Ser Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser
225 230 235 240
Ala Asp Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu
245 250 255
Thr Glu Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly
260 265 270
Trp Gly Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu
275 280 285
His Asn Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg
290 295 300
Ser Arg Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro
305 310 315 320
His Pro Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val
325 330 335
Leu Phe Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala
340 345 350
Leu Glu Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro
355 360 365
Gly Gly Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser
370 375 380
Gln Trp Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg
385 390 395 400
Asp Lys Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu
405 410 415
Thr Leu Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu
420 425 430
Ala Gly Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser
435 440 445
Leu Gly Ala Ala Pro Ala Ala Ala Pro Ala Lys Gln Glu Ala Ala Ala
450 455 460
Pro Ala Pro Ala Ala Lys Ala Glu Ala Pro Ala Ala Ala Pro Ala Ala
465 470 475 480
Lys Ala Thr Pro Gln Gln Leu Glu Asp Lys Val Glu Glu Leu Leu Ser
485 490 495
Lys Asn Tyr His Leu Glu Asn Glu Val Ala Arg Leu Lys Lys Leu Val
500 505 510
Gly
<210> 116
<211> 1569
<212> DNA
<213> Artificial Sequence
<400> 116
atgagggcca agcagctgga ggacaaggtc gaggagctgc tgagcaagaa ctaccacctg 60
gagaacgagg tcgcccgcct gaagaagctg gtgggcaccc gcagcgcagc cgaagccgct 120
gcgaaggagg cagctgcgaa agaagcggct gcaaaagaag cggcagctaa ggctttgaat 180
accccgcaat cggctttcgc tgcccaatcg gaaccggaac tgaaactgga aagtgtggtt 240
attgtgtctc gtcatggcgt tcgcgctccg accaaattta cgcagctgat gcaagatgtc 300
accccggacg ccttctatac gtggccggtg aagctgggtg aactgacccc gcgtggcggt 360
gaactgatcg cctatctggg tcactactgg cgtcagcgcc tggtggcaga tggtctgctg 420
ccgaaaaagg gctgcccgca gagcggtcaa gttgcaatta tcgctgatgt cgacgaacgt 480
acccgcaaaa cgggtgaagc atttgcggcc ggtctggcac cggattgcgc cattaccgtt 540
catacgcagg cagataccag ctctccggac ccgctgttca acccgctgaa aaccggcgtc 600
tgtcagctgg atgtcgcgca agtgacggac gccattctgg aacgtgcagg cggttccatc 660
gctgatttta ccggtcacta ccagacggca ttccgtgaac tggaacgcgt tctgaacttt 720
ccgcagtcaa atctggcgct gaaacgcgaa aagcaggatg aaagtgcgtc cctgacccaa 780
gccctgccga gtgaactgaa agtctccgcc gacaatgtgt cactgaccgg cgcatggtca 840
ctggcttcga tgctgacgga aatttttctg ctgcagcaag cacagggtat gccggaaccg 900
ggttggggtc gtatcaccga ttcgcatcag tggaacacgc tgctgagcct gcacaatgcg 960
cagttcgacc tgctgcaacg taccccggaa gtggcacgtt cgcgcgccac gccgctgctg 1020
gatctgatta aaaccgctct gacgccgcat ccgccgcaga agcaagcgta tggcgtgacc 1080
ctgccgacga gcgttctgtt tatcgcgggt cacgacacca acctggcaaa tctgggcggt 1140
gctctggaac tgcagtggac cctgccgggt caaccggata acacgccgcc gggcggtgaa 1200
ctggttttcg aacgttggcg tcgcctgagc gacaattctc agtggatcca agttagcctg 1260
gtctttcaga ccctgcagca aatgcgcgat aaaaccccgc tgttcctgaa cacgccgccg 1320
ggcgaagtga agctgaccct ggcgggttgc gaagaacgta acgcccaggg catgtgttct 1380
ctggcaggtt ttacccagat tgttaatgaa gcacgcatcc cggcttgtag tctggggggc 1440
gcagaagcag ctgccaaaga ggcggccgca aaggtcaatc tgcaattgga agataaagtg 1500
gaagagctcc tgtccaaaaa ttatcatctg gaaaatgagg tggcccgctt gaagaaactc 1560
gtgggataa 1569
<210> 117
<211> 522
<212> PRT
<213> Artificial Sequence
<400> 117
Met Arg Ala Lys Gln Leu Glu Asp Lys Val Glu Glu Leu Leu Ser Lys
1 5 10 15
Asn Tyr His Leu Glu Asn Glu Val Ala Arg Leu Lys Lys Leu Val Gly
20 25 30
Thr Arg Ser Ala Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Glu
35 40 45
Ala Ala Ala Lys Glu Ala Ala Ala Lys Ala Leu Asn Thr Pro Gln Ser
50 55 60
Ala Phe Ala Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val Val
65 70 75 80
Ile Val Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe Thr Gln Leu
85 90 95
Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro Val Lys Leu
100 105 110
Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu Gly His
115 120 125
Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys Lys Gly
130 135 140
Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp Glu Arg
145 150 155 160
Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro Asp Cys
165 170 175
Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro Asp Pro Leu
180 185 190
Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val Ala Gln Val
195 200 205
Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp Phe Thr
210 215 220
Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu Asn Phe
225 230 235 240
Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp Glu Ser Ala
245 250 255
Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala Asp Asn
260 265 270
Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu Thr Glu Ile
275 280 285
Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp Gly Arg
290 295 300
Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu His Asn Ala
305 310 315 320
Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser Arg Ala
325 330 335
Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His Pro Pro
340 345 350
Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu Phe Ile
355 360 365
Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu Glu Leu
370 375 380
Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly Glu
385 390 395 400
Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln Trp Ile
405 410 415
Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp Lys Thr
420 425 430
Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr Leu Ala
435 440 445
Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala Gly Phe
450 455 460
Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu Gly Gly
465 470 475 480
Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Val Asn Leu Gln Leu
485 490 495
Glu Asp Lys Val Glu Glu Leu Leu Ser Lys Asn Tyr His Leu Glu Asn
500 505 510
Glu Val Ala Arg Leu Lys Lys Leu Val Gly
515 520
<210> 118
<211> 1659
<212> DNA
<213> Artificial Sequence
<400> 118
atggcccaca tcgtgatggt ggacgcctac aagccgacga agggttcagg gggttccggt 60
gcccaatcgg aaccggaact gaaactggaa agtgtggtta ttgtgtctcg tcatggcgtt 120
cgcgctccga ccaaatttac gcagctgatg caagatgtca ccccggacgc cttctatacg 180
tggccggtga agctgggtga actgaccccg cgtggcggtg aactgatcgc ctatctgggt 240
cactactggc gtcagcgcct ggtggcagat ggtctgctgc cgaaaaaggg ctgcccgcag 300
agcggtcaag ttgcaattat cgctgatgtc gacgaacgta cccgcaaaac gggtgaagca 360
tttgcggccg gtctggcacc ggattgcgcc attaccgttc atacgcaggc agataccagc 420
tctccggacc cgctgttcaa cccgctgaaa accggcgtct gtcagctgga tgtcgcgcaa 480
gtgacggacg ccattctgga acgtgcaggc ggttccatcg ctgattttac cggtcactac 540
cagacggcat tccgtgaact ggaacgcgtt ctgaactttc cgcagtcaaa tctggcgctg 600
aaacgcgaaa agcaggatga aagtgcgtcc ctgacccaag ccctgccgag tgaactgaaa 660
gtctccgccg acaatgtgtc actgaccggc gcatggtcac tggcttcgat gctgacggaa 720
atttttctgc tgcagcaagc acagggtatg ccggaaccgg gttggggtcg tatcaccgat 780
tcgcatcagt ggaacacgct gctgagcctg cacaatgcgc agttcgacct gctgcaacgt 840
accccggaag tggcacgttc gcgcgccacg ccgctgctgg atctgattaa aaccgctctg 900
acgccgcatc cgccgcagaa gcaagcgtat ggcgtgaccc tgccgacgag cgttctgttt 960
atcgcgggtc acgacaccaa cctggcaaat ctgggcggtg ctctggaact gcagtggacc 1020
ctgccgggtc aaccggataa cacgccgccg ggcggtgaac tggttttcga acgttggcgt 1080
cgcctgagcg acaattctca gtggatccaa gttagcctgg tctttcagac cctgcagcaa 1140
atgcgcgata aaaccccgct gttcctgaac acgccgccgg gcgaagtgaa gctgaccctg 1200
gcgggttgcg aagaacgtaa cgcccagggc atgtgttctc tggcaggttt tacccagatt 1260
gttaatgaag cacgcatccc ggcttgtagt ctggggagtg gtggcagcgg aggcgctatg 1320
gttgatacct tatcaggttt atcaagtgag caaggtcagt ccggtgatat gacaattgaa 1380
gaagatagtg ctacccatat taaattctca aaacgtgatg aggacggcaa agagttagct 1440
ggtgcaacta tggagttgcg tgattcatct ggtaaaacta ttagtacatg gatttcagat 1500
ggacaagtga aagatttcta cctgtatcca ggaaaatata catttgtcga aaccgcagca 1560
ccagacggtt atgaggtagc aactgctatt acctttacag ttaatgagca aggtcaggtt 1620
actgtaaatg gcaaagcaac taaaggtgac gctcatatt 1659
<210> 119
<211> 553
<212> PRT
<213> Artificial Sequence
<400> 119
Met Ala His Ile Val Met Val Asp Ala Tyr Lys Pro Thr Lys Gly Ser
1 5 10 15
Gly Gly Ser Gly Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val
20 25 30
Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe Thr Gln
35 40 45
Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro Val Lys
50 55 60
Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu Gly
65 70 75 80
His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys Lys
85 90 95
Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp Glu
100 105 110
Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro Asp
115 120 125
Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro Asp Pro
130 135 140
Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val Ala Gln
145 150 155 160
Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp Phe
165 170 175
Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu Asn
180 185 190
Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp Glu Ser
195 200 205
Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala Asp
210 215 220
Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu Thr Glu
225 230 235 240
Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp Gly
245 250 255
Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu His Asn
260 265 270
Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser Arg
275 280 285
Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His Pro
290 295 300
Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu Phe
305 310 315 320
Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu Glu
325 330 335
Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly
340 345 350
Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln Trp
355 360 365
Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp Lys
370 375 380
Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr Leu
385 390 395 400
Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala Gly
405 410 415
Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu Gly
420 425 430
Ser Gly Gly Ser Gly Gly Ala Met Val Asp Thr Leu Ser Gly Leu Ser
435 440 445
Ser Glu Gln Gly Gln Ser Gly Asp Met Thr Ile Glu Glu Asp Ser Ala
450 455 460
Thr His Ile Lys Phe Ser Lys Arg Asp Glu Asp Gly Lys Glu Leu Ala
465 470 475 480
Gly Ala Thr Met Glu Leu Arg Asp Ser Ser Gly Lys Thr Ile Ser Thr
485 490 495
Trp Ile Ser Asp Gly Gln Val Lys Asp Phe Tyr Leu Tyr Pro Gly Lys
500 505 510
Tyr Thr Phe Val Glu Thr Ala Ala Pro Asp Gly Tyr Glu Val Ala Thr
515 520 525
Ala Ile Thr Phe Thr Val Asn Glu Gln Gly Gln Val Thr Val Asn Gly
530 535 540
Lys Ala Thr Lys Gly Asp Ala His Ile
545 550
<210> 120
<211> 48
<212> DNA
<213> Artificial Sequence
<400> 120
agcggcggcg gcagcggcgg cggcagcacc ccgcagagcg ccttcgcc 48
<210> 121
<211> 33
<212> DNA
<213> Artificial Sequence
<400> 121
ggcggcggca gcggcggcgg cagcggcggc ggc 33
<210> 122
<211> 60
<212> DNA
<213> Artificial Sequence
<400> 122
agcggcggca gcagcggcag caccaccacc accaggatca ccccgcagag cgccttcgcc 60
<210> 123
<211> 36
<212> DNA
<213> Artificial Sequence
<400> 123
cagaacacct tcagccaggg cagcagcagc ggcagc 36
<210> 124
<211> 12
<212> DNA
<213> Artificial Sequence
<400> 124
agcgccttcg cc 12
<210> 125
<211> 108
<212> DNA
<213> Artificial Sequence
<400> 125
ggcgccgccc cggccgccgc cccggccaag caggaggccg ccgccccggc cccggccgcc 60
aaggccgagg ccccggccgc cgccccggcc gccaaggcca ccccgcag 108
<210> 126
<211> 99
<212> DNA
<213> Artificial Sequence
<400> 126
agcgccgccg aggccgccgc caaggaggcc gccgccaagg aggccgccgc caaggaggcc 60
gccgccaagg ccctgaacac cccgcagagc gccttcgcc 99
<210> 127
<211> 48
<212> DNA
<213> Artificial Sequence
<400> 127
ggcggcgccg aggccgccgc caaggaggcc gccgccaagg tgaacctg 48
<210> 128
<211> 3274
<212> DNA
<213> Artificial Sequence
<400> 128
ggtaccaaag taatcatatt attttatgtg tgaatcttct ttactttttc atttgattat 60
gattatgaag gtatgacctt cataaccttc gtccgaaatc cattatatcc aaaggaaaat 120
aatgcttcga aggacgaagg attttgatat ttaacatttt atgttgcctt gttcttaatt 180
catagcattt gagaacaagt ccccaacacc aatctttatc tttactatat taaagcacca 240
gttcaacgat cgtctcgtgt caatattatt aaaaaactcc tacatttctt tataatcaac 300
ccgcactctt ataatctctt ctcttactac tataataaga gagtttatgt acaaaataag 360
gtgaaattat gtataagtgt tctggacctt ggttgttggc tcatattcac acaacctaat 420
caatagaaaa catatgtttt attaaaacaa aatttatcat atatatatat atatatatat 480
atatatatat atatatatat aatataaacc gtagcaatgc acaggcatat gactagtggc 540
aacttaatac catgtgtgta ttaagatgaa taagaggtat ccaaataaat aacttgttcg 600
cttacgtctg gatcgaaagg ggttggaaac gattaaatct cttcctagtc aaaattaaat 660
agaaggagat ttaatcgatt tctcccaatc cccttcgatc caggtgcaac cgaataagtc 720
cttaaatgtt gaggaacacg aaacaaccat gcattggcat gtaaagctcc aagaattcgt 780
tgtatcctta acaactcaca gaacatcaac caaaattgca cgtcaagggt attgggtaag 840
aaacaatcaa acaaatcctc tctgtgtgca aagaaacacg gtgagtcatg ccgagatcat 900
actcatctga tatacatgct tacagctcac aagacattac aaacaactca tattgcatta 960
caaagatcgt ttcatgaaaa ataaaatagg ccggaacagg acaaaaatcc ttgacgtgta 1020
aagtaaattt acaacaaaaa aaaagccata tgtcaagcta aatctaattc gttttacgta 1080
gatcaacaac ctgtagaagg caacaaaact gagccacgca gaagtacaga atgattccag 1140
atgaaccatc gacgtgctac gtaaagagag tgacgagtca tatacatttg gcaagaaacc 1200
atgaagctgc ctacagccgt ctcggtggca taagaacaca agaaattgtg ttaattaatc 1260
aaagctataa ataacgctcg catgcctgtg cacttctcca tcaccaccac tgggtcttca 1320
gaccattagc tttatctact ccagagcgca gaagaacccg atcgacaccg gatccaccat 1380
gatgctgaag aagatcctga agatcgagga gctggacgag agggagctga tcgacatcga 1440
ggtgagcggc aaccacctgt tctacgccaa cgacatcctg acccacaaca gcgcccagtc 1500
cgagccggag ctgaagctgg agtccgtggt gatcgtgtcg cgccacgggg tgcgcgcccc 1560
gaccaagttc acgcagctca tgcaggacgt gaccccggac gccttctaca cctggccggt 1620
gaagctcggc gagctgaccc cgcgcggcgg cgagctgatc gcctacctcg gccactactg 1680
gcgccagcgc ctcgtggccg acggcctcct cccgaagaag ggctgcccgc agtccggcca 1740
ggtggcgatc atcgccgacg tggacgagcg cacccgcaag acgggcgagg ccttcgccgc 1800
cggcctcgcc ccggactgcg ccatcaccgt gcacacccag gccgacacct cctccccgga 1860
cccgctcttc aacccgctca agaccggcgt gtgccagctc gacgtggccc aggtgaccga 1920
cgccatcctg gagcgcgccg gcggctccat cgccgacttc accggccact accagaccgc 1980
cttccgcgag ctggagcgcg tgctcaactt cccgcagtcg aacctcgccc tcaagcgcga 2040
gaagcaggac gagtccgcct ccctcaccca ggccctcccg tccgagctga aggtgtccgc 2100
cgacaacgtg tccctcaccg gcgcctggtc cctcgcctcc atgctcaccg aaatcttcct 2160
cctccagcag gcccagggca tgccggagcc gggctggggc cgcatcaccg actcccacca 2220
gtggaacacc ctcctctccc tccacaacgc ccagttcgac ctcctccagc gcaccccgga 2280
ggtggcccgc tcccgcgcca ccccgctcct cgacctcatc aagaccgccc tcaccccgca 2340
cccgccgcag aagcaggcct acggcgtgac cctcccgacc tcggtgctct tcatcgccgg 2400
ccacgacacc aacctcgcca acctcggcgg cgccctggag ctgcagtgga ccctcccggg 2460
ccagccggac aacaccccgc cgggcggcga gctggtgttc gagcgctggc gccgcctctc 2520
cgacaactcc cagtggattc aggtgtccct cgtgttccag accctccagc agatgcgcga 2580
caagaccccg ctcttcctca acaccccgcc gggcgaggtg aagctcaccc tggccggctg 2640
cgaggagcgc aacgcgcagg gcatgtgctc cctcgccggc ttcacccaga tcgtgaacga 2700
ggcccgcatc ccggcctgct ccctctgcct ggacctgaag acccaggtgc agaccccgca 2760
gggcatgaag gagatcagca acatccaggt gggcgacctg gtgctgagca acaccggcta 2820
caacgaggtg ctgaacgtgt tcccgaagag caagaagaag agctacaaga tcaccctgga 2880
ggacggcaag gagatcatct gcagcgagga gcacctgttc ccgacccaga ccggcgagat 2940
gaacatcagc ggcggcctga aggagggcat gtgcctgtac gtgaaggagt gacctaggtc 3000
cccgaatttc cccgatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt 3060
tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc atgtaataat 3120
taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt 3180
atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg 3240
cgcggtgtca tctatgttac tagatcggga attg 3274
<210> 129
<211> 3364
<212> DNA
<213> Artificial Sequence
<400> 129
ggtaccaaag taatcatatt attttatgtg tgaatcttct ttactttttc atttgattat 60
gattatgaag gtatgacctt cataaccttc gtccgaaatc cattatatcc aaaggaaaat 120
aatgcttcga aggacgaagg attttgatat ttaacatttt atgttgcctt gttcttaatt 180
catagcattt gagaacaagt ccccaacacc aatctttatc tttactatat taaagcacca 240
gttcaacgat cgtctcgtgt caatattatt aaaaaactcc tacatttctt tataatcaac 300
ccgcactctt ataatctctt ctcttactac tataataaga gagtttatgt acaaaataag 360
gtgaaattat gtataagtgt tctggacctt ggttgttggc tcatattcac acaacctaat 420
caatagaaaa catatgtttt attaaaacaa aatttatcat atatatatat atatatatat 480
atatatatat atatatatat aatataaacc gtagcaatgc acaggcatat gactagtggc 540
aacttaatac catgtgtgta ttaagatgaa taagaggtat ccaaataaat aacttgttcg 600
cttacgtctg gatcgaaagg ggttggaaac gattaaatct cttcctagtc aaaattaaat 660
agaaggagat ttaatcgatt tctcccaatc cccttcgatc caggtgcaac cgaataagtc 720
cttaaatgtt gaggaacacg aaacaaccat gcattggcat gtaaagctcc aagaattcgt 780
tgtatcctta acaactcaca gaacatcaac caaaattgca cgtcaagggt attgggtaag 840
aaacaatcaa acaaatcctc tctgtgtgca aagaaacacg gtgagtcatg ccgagatcat 900
actcatctga tatacatgct tacagctcac aagacattac aaacaactca tattgcatta 960
caaagatcgt ttcatgaaaa ataaaatagg ccggaacagg acaaaaatcc ttgacgtgta 1020
aagtaaattt acaacaaaaa aaaagccata tgtcaagcta aatctaattc gttttacgta 1080
gatcaacaac ctgtagaagg caacaaaact gagccacgca gaagtacaga atgattccag 1140
atgaaccatc gacgtgctac gtaaagagag tgacgagtca tatacatttg gcaagaaacc 1200
atgaagctgc ctacagccgt ctcggtggca taagaacaca agaaattgtg ttaattaatc 1260
aaagctataa ataacgctcg catgcctgtg cacttctcca tcaccaccac tgggtcttca 1320
gaccattagc tttatctact ccagagcgca gaagaacccg atcgacaccg gatccaccat 1380
gagggtgttg ctcgttgccc tcgctctcct ggctctcgct gcgagcgcca ccagcatgat 1440
gctgaagaag atcctgaaga tcgaggagct ggacgagagg gagctgatcg acatcgaggt 1500
gagcggcaac cacctgttct acgccaacga catcctgacc cacaacagcg ctgcgcagtc 1560
cgagccggag ctgaagctgg agtccgtggt gatcgtgtcg cgccacgggg tgcgcgcccc 1620
gaccaagttc acgcagctca tgcaggacgt gaccccggac gccttctaca cctggccggt 1680
gaagctcggc gagctgaccc cgcgcggcgg cgagctgatc gcctacctcg gccactactg 1740
gcgccagcgc ctcgtggccg acggcctcct cccgaagaag ggctgcccgc agtccggcca 1800
ggtggcgatc atcgccgacg tggacgagcg cacccgcaag acgggcgagg ccttcgccgc 1860
cggcctcgcc ccggactgcg ccatcaccgt gcacacccag gccgacacct cctccccgga 1920
cccgctcttc aacccgctca agaccggcgt gtgccagctc gacgtggccc aggtgaccga 1980
cgccatcctg gagcgcgccg gcggctccat cgccgacttc accggccact accagaccgc 2040
cttccgcgag ctggagcgcg tgctcaactt cccgcagtcg aacctcgccc tcaagcgcga 2100
gaagcaggac gagtccgcct ccctcaccca ggccctcccg tccgagctga aggtgtccgc 2160
cgacaacgtg tccctcaccg gcgcctggtc cctcgcctcc atgctcaccg aaatcttcct 2220
cctccagcag gcccagggca tgccggagcc gggctggggc cgcatcaccg actcccacca 2280
gtggaacacc ctcctctccc tccacaacgc ccagttcgac ctcctccagc gcaccccgga 2340
ggtggcccgc tcccgcgcca ccccgctcct cgacctcatc aagaccgccc tcaccccgca 2400
cccgccgcag aagcaggcct acggcgtgac cctcccgacc tcggtgctct tcatcgccgg 2460
ccacgacacc aacctcgcca acctcggcgg cgccctggag ctgcagtgga ccctcccggg 2520
ccagccggac aacaccccgc cgggcggcga gctggtgttc gagcgctggc gccgcctctc 2580
cgacaactcc cagtggattc aggtgtccct cgtgttccag accctccagc agatgcgcga 2640
caagaccccg ctcttcctca acaccccgcc gggcgaggtg aagctcaccc tggccggctg 2700
cgaggagcgc aacgcgcagg gcatgtgctc cctcgccggc ttcacccaga tcgtgaacga 2760
ggcccgcatc ccggcctgct ccctctgcct ggacctgaag acccaggtgc agaccccgca 2820
gggcatgaag gagatcagca acatccaggt gggcgacctg gtgctgagca acaccggcta 2880
caacgaggtg ctgaacgtgt tcccgaagag caagaagaag agctacaaga tcaccctgga 2940
ggacggcaag gagatcatct gcagcgagga gcacctgttc ccgacccaga ccggcgagat 3000
gaacatcagc ggcggcctga aggagggcat gtgcctgtac gtgaaggagg acccgaacgg 3060
ctccgagaag gacgagctgt gacctaggtc cccgaatttc cccgatcgtt caaacatttg 3120
gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt 3180
tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag 3240
atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat 3300
atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac tagatcggga 3360
attg 3364
<210> 130
<211> 3379
<212> DNA
<213> Artificial Sequence
<400> 130
ggtaccaaag taatcatatt attttatgtg tgaatcttct ttactttttc atttgattat 60
gattatgaag gtatgacctt cataaccttc gtccgaaatc cattatatcc aaaggaaaat 120
aatgcttcga aggacgaagg attttgatat ttaacatttt atgttgcctt gttcttaatt 180
catagcattt gagaacaagt ccccaacacc aatctttatc tttactatat taaagcacca 240
gttcaacgat cgtctcgtgt caatattatt aaaaaactcc tacatttctt tataatcaac 300
ccgcactctt ataatctctt ctcttactac tataataaga gagtttatgt acaaaataag 360
gtgaaattat gtataagtgt tctggacctt ggttgttggc tcatattcac acaacctaat 420
caatagaaaa catatgtttt attaaaacaa aatttatcat atatatatat atatatatat 480
atatatatat atatatatat aatataaacc gtagcaatgc acaggcatat gactagtggc 540
aacttaatac catgtgtgta ttaagatgaa taagaggtat ccaaataaat aacttgttcg 600
cttacgtctg gatcgaaagg ggttggaaac gattaaatct cttcctagtc aaaattaaat 660
agaaggagat ttaatcgatt tctcccaatc cccttcgatc caggtgcaac cgaataagtc 720
cttaaatgtt gaggaacacg aaacaaccat gcattggcat gtaaagctcc aagaattcgt 780
tgtatcctta acaactcaca gaacatcaac caaaattgca cgtcaagggt attgggtaag 840
aaacaatcaa acaaatcctc tctgtgtgca aagaaacacg gtgagtcatg ccgagatcat 900
actcatctga tatacatgct tacagctcac aagacattac aaacaactca tattgcatta 960
caaagatcgt ttcatgaaaa ataaaatagg ccggaacagg acaaaaatcc ttgacgtgta 1020
aagtaaattt acaacaaaaa aaaagccata tgtcaagcta aatctaattc gttttacgta 1080
gatcaacaac ctgtagaagg caacaaaact gagccacgca gaagtacaga atgattccag 1140
atgaaccatc gacgtgctac gtaaagagag tgacgagtca tatacatttg gcaagaaacc 1200
atgaagctgc ctacagccgt ctcggtggca taagaacaca agaaattgtg ttaattaatc 1260
aaagctataa ataacgctcg catgcctgtg cacttctcca tcaccaccac tgggtcttca 1320
gaccattagc tttatctact ccagagcgca gaagaacccg atcgacaccg gatccaccat 1380
ggttaaggtg attggaagac gttctcttgg tgttcaaagg atcttcgata tcggattgcc 1440
acaagaccac aactttcttc tcgctaatgg tgccatcgct gccaatagcg ctgcgcagtc 1500
cgagccggag ctgaagctgg agtccgtggt gatcgtgtcg cgccacgggg tgcgcgcccc 1560
gaccaagttc acgcagctca tgcaggacgt gaccccggac gccttctaca cctggccggt 1620
gaagctcggc gagctgaccc cgcgcggcgg cgagctgatc gcctacctcg gccactactg 1680
gcgccagcgc ctcgtggccg acggcctcct cccgaagaag ggctgcccgc agtccggcca 1740
ggtggcgatc atcgccgacg tggacgagcg cacccgcaag acgggcgagg ccttcgccgc 1800
cggcctcgcc ccggactgcg ccatcaccgt gcacacccag gccgacacct cctccccgga 1860
cccgctcttc aacccgctca agaccggcgt gtgccagctc gacgtggccc aggtgaccga 1920
cgccatcctg gagcgcgccg gcggctccat cgccgacttc accggccact accagaccgc 1980
cttccgcgag ctggagcgcg tgctcaactt cccgcagtcg aacctcgccc tcaagcgcga 2040
gaagcaggac gagtccgcct ccctcaccca ggccctcccg tccgagctga aggtgtccgc 2100
cgacaacgtg tccctcaccg gcgcctggtc cctcgcctcc atgctcaccg aaatcttcct 2160
cctccagcag gcccagggca tgccggagcc gggctggggc cgcatcaccg actcccacca 2220
gtggaacacc ctcctctccc tccacaacgc ccagttcgac ctcctccagc gcaccccgga 2280
ggtggcccgc tcccgcgcca ccccgctcct cgacctcatc aagaccgccc tcaccccgca 2340
cccgccgcag aagcaggcct acggcgtgac cctcccgacc tcggtgctct tcatcgccgg 2400
ccacgacacc aacctcgcca acctcggcgg cgccctggag ctgcagtgga ccctcccggg 2460
ccagccggac aacaccccgc cgggcggcga gctggtgttc gagcgctggc gccgcctctc 2520
cgacaactcc cagtggattc aggtgtccct cgtgttccag accctccagc agatgcgcga 2580
caagaccccg ctcttcctca acaccccgcc gggcgaggtg aagctcaccc tggccggctg 2640
cgaggagcgc aacgcgcagg gcatgtgctc cctcgccggc ttcacccaga tcgtgaacga 2700
ggcccgcatc ccggcctgct ccctctgcct ttctttcgga actgagatcc ttaccgttga 2760
gtacggacca cttcctattg gtaagatcgt ttctgaggaa attaactgct cagtgtactc 2820
tgttgatcca gaaggaagag tttacactca ggctatcgca caatggcacg ataggggtga 2880
acaagaggtt ctggagtacg agcttgaaga tggatccgtt attcgtgcta cctctgacca 2940
tagattcttg actacagatt atcagcttct cgctatcgag gaaatctttg ctaggcaact 3000
tgatctcctt actttggaga acatcaagca gacagaagag gctcttgaca accacagact 3060
tccattccct ttgctcgatg ctggaaccat caagtaacct aggtccccga atttccccga 3120
tcgttcaaac atttggcaat aaagtttctt aagattgaat cctgttgccg gtcttgcgat 3180
gattatcata taatttctgt tgaattacgt taagcatgta ataattaaca tgtaatgcat 3240
gacgttattt atgagatggg tttttatgat tagagtcccg caattataca tttaatacgc 3300
gatagaaaac aaaatatagc gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat 3360
gttactagat cgggaattg 3379
<210> 131
<211> 3466
<212> DNA
<213> Artificial Sequence
<400> 131
ggtaccaaag taatcatatt attttatgtg tgaatcttct ttactttttc atttgattat 60
gattatgaag gtatgacctt cataaccttc gtccgaaatc cattatatcc aaaggaaaat 120
aatgcttcga aggacgaagg attttgatat ttaacatttt atgttgcctt gttcttaatt 180
catagcattt gagaacaagt ccccaacacc aatctttatc tttactatat taaagcacca 240
gttcaacgat cgtctcgtgt caatattatt aaaaaactcc tacatttctt tataatcaac 300
ccgcactctt ataatctctt ctcttactac tataataaga gagtttatgt acaaaataag 360
gtgaaattat gtataagtgt tctggacctt ggttgttggc tcatattcac acaacctaat 420
caatagaaaa catatgtttt attaaaacaa aatttatcat atatatatat atatatatat 480
atatatatat atatatatat aatataaacc gtagcaatgc acaggcatat gactagtggc 540
aacttaatac catgtgtgta ttaagatgaa taagaggtat ccaaataaat aacttgttcg 600
cttacgtctg gatcgaaagg ggttggaaac gattaaatct cttcctagtc aaaattaaat 660
agaaggagat ttaatcgatt tctcccaatc cccttcgatc caggtgcaac cgaataagtc 720
cttaaatgtt gaggaacacg aaacaaccat gcattggcat gtaaagctcc aagaattcgt 780
tgtatcctta acaactcaca gaacatcaac caaaattgca cgtcaagggt attgggtaag 840
aaacaatcaa acaaatcctc tctgtgtgca aagaaacacg gtgagtcatg ccgagatcat 900
actcatctga tatacatgct tacagctcac aagacattac aaacaactca tattgcatta 960
caaagatcgt ttcatgaaaa ataaaatagg ccggaacagg acaaaaatcc ttgacgtgta 1020
aagtaaattt acaacaaaaa aaaagccata tgtcaagcta aatctaattc gttttacgta 1080
gatcaacaac ctgtagaagg caacaaaact gagccacgca gaagtacaga atgattccag 1140
atgaaccatc gacgtgctac gtaaagagag tgacgagtca tatacatttg gcaagaaacc 1200
atgaagctgc ctacagccgt ctcggtggca taagaacaca agaaattgtg ttaattaatc 1260
aaagctataa ataacgctcg catgcctgtg cacttctcca tcaccaccac tgggtcttca 1320
gaccattagc tttatctact ccagagcgca gaagaacccg atcgacaccg gatccaccat 1380
gagggtgttg ctcgttgccc tcgctctcct ggctctcgct gcgagcgcca ccagcatggt 1440
taaggtgatt ggaagacgtt ctcttggtgt tcaaaggatc ttcgatatcg gattgccaca 1500
agaccacaac tttcttctcg ctaatggtgc catcgctgcc aatagcgctg cgcagtccga 1560
gccggagctg aagctggagt ccgtggtgat cgtgtcgcgc cacggggtgc gcgccccgac 1620
caagttcacg cagctcatgc aggacgtgac cccggacgcc ttctacacct ggccggtgaa 1680
gctcggcgag ctgaccccgc gcggcggcga gctgatcgcc tacctcggcc actactggcg 1740
ccagcgcctc gtggccgacg gcctcctccc gaagaagggc tgcccgcagt ccggccaggt 1800
ggcgatcatc gccgacgtgg acgagcgcac ccgcaagacg ggcgaggcct tcgccgccgg 1860
cctcgccccg gactgcgcca tcaccgtgca cacccaggcc gacacctcct ccccggaccc 1920
gctcttcaac ccgctcaaga ccggcgtgtg ccagctcgac gtggcccagg tgaccgacgc 1980
catcctggag cgcgccggcg gctccatcgc cgacttcacc ggccactacc agaccgcctt 2040
ccgcgagctg gagcgcgtgc tcaacttccc gcagtcgaac ctcgccctca agcgcgagaa 2100
gcaggacgag tccgcctccc tcacccaggc cctcccgtcc gagctgaagg tgtccgccga 2160
caacgtgtcc ctcaccggcg cctggtccct cgcctccatg ctcaccgaaa tcttcctcct 2220
ccagcaggcc cagggcatgc cggagccggg ctggggccgc atcaccgact cccaccagtg 2280
gaacaccctc ctctccctcc acaacgccca gttcgacctc ctccagcgca ccccggaggt 2340
ggcccgctcc cgcgccaccc cgctcctcga cctcatcaag accgccctca ccccgcaccc 2400
gccgcagaag caggcctacg gcgtgaccct cccgacctcg gtgctcttca tcgccggcca 2460
cgacaccaac ctcgccaacc tcggcggcgc cctggagctg cagtggaccc tcccgggcca 2520
gccggacaac accccgccgg gcggcgagct ggtgttcgag cgctggcgcc gcctctccga 2580
caactcccag tggattcagg tgtccctcgt gttccagacc ctccagcaga tgcgcgacaa 2640
gaccccgctc ttcctcaaca ccccgccggg cgaggtgaag ctcaccctgg ccggctgcga 2700
ggagcgcaac gcgcagggca tgtgctccct cgccggcttc acccagatcg tgaacgaggc 2760
ccgcatcccg gcctgctccc tctgcctttc tttcggaact gagatcctta ccgttgagta 2820
cggaccactt cctattggta agatcgtttc tgaggaaatt aactgctcag tgtactctgt 2880
tgatccagaa ggaagagttt acactcaggc tatcgcacaa tggcacgata ggggtgaaca 2940
agaggttctg gagtacgagc ttgaagatgg atccgttatt cgtgctacct ctgaccatag 3000
attcttgact acagattatc agcttctcgc tatcgaggaa atctttgcta ggcaacttga 3060
tctccttact ttggagaaca tcaagcagac agaagaggct cttgacaacc acagacttcc 3120
attccctttg ctcgatgctg gaaccatcaa ggacccgaac ggctccgaga aggacgagct 3180
gtaacctagg tccccgaatt tccccgatcg ttcaaacatt tggcaataaa gtttcttaag 3240
attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga attacgttaa 3300
gcatgtaata attaacatgt aatgcatgac gttatttatg agatgggttt ttatgattag 3360
agtcccgcaa ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga 3420
taaattatcg cgcgcggtgt catctatgtt actagatcgg gaattg 3466
<210> 132
<211> 3457
<212> DNA
<213> Artificial Sequence
<400> 132
ggtaccaaag taatcatatt attttatgtg tgaatcttct ttactttttc atttgattat 60
gattatgaag gtatgacctt cataaccttc gtccgaaatc cattatatcc aaaggaaaat 120
aatgcttcga aggacgaagg attttgatat ttaacatttt atgttgcctt gttcttaatt 180
catagcattt gagaacaagt ccccaacacc aatctttatc tttactatat taaagcacca 240
gttcaacgat cgtctcgtgt caatattatt aaaaaactcc tacatttctt tataatcaac 300
ccgcactctt ataatctctt ctcttactac tataataaga gagtttatgt acaaaataag 360
gtgaaattat gtataagtgt tctggacctt ggttgttggc tcatattcac acaacctaat 420
caatagaaaa catatgtttt attaaaacaa aatttatcat atatatatat atatatatat 480
atatatatat atatatatat aatataaacc gtagcaatgc acaggcatat gactagtggc 540
aacttaatac catgtgtgta ttaagatgaa taagaggtat ccaaataaat aacttgttcg 600
cttacgtctg gatcgaaagg ggttggaaac gattaaatct cttcctagtc aaaattaaat 660
agaaggagat ttaatcgatt tctcccaatc cccttcgatc caggtgcaac cgaataagtc 720
cttaaatgtt gaggaacacg aaacaaccat gcattggcat gtaaagctcc aagaattcgt 780
tgtatcctta acaactcaca gaacatcaac caaaattgca cgtcaagggt attgggtaag 840
aaacaatcaa acaaatcctc tctgtgtgca aagaaacacg gtgagtcatg ccgagatcat 900
actcatctga tatacatgct tacagctcac aagacattac aaacaactca tattgcatta 960
caaagatcgt ttcatgaaaa ataaaatagg ccggaacagg acaaaaatcc ttgacgtgta 1020
aagtaaattt acaacaaaaa aaaagccata tgtcaagcta aatctaattc gttttacgta 1080
gatcaacaac ctgtagaagg caacaaaact gagccacgca gaagtacaga atgattccag 1140
atgaaccatc gacgtgctac gtaaagagag tgacgagtca tatacatttg gcaagaaacc 1200
atgaagctgc ctacagccgt ctcggtggca taagaacaca agaaattgtg ttaattaatc 1260
aaagctataa ataacgctcg catgcctgtg cacttctcca tcaccaccac tgggtcttca 1320
gaccattagc tttatctact ccagagcgca gaagaacccg atcgacaccg gatccaccat 1380
ggttaaggtg attggaagac gttctcttgg tgttcaaagg atcttcgata tcggattgcc 1440
acaagaccac aactttcttc tcgctaatgg tgccatcgct gccaatagcg gcggcggcag 1500
cggcggcggc agcaccccgc agagcgcctt cgccgctgcg cagtccgagc cggagctgaa 1560
gctggagtcc gtggtgatcg tgtcgcgcca cggggtgcgc gccccgacca agttcacgca 1620
gctcatgcag gacgtgaccc cggacgcctt ctacacctgg ccggtgaagc tcggcgagct 1680
gaccccgcgc ggcggcgagc tgatcgccta cctcggccac tactggcgcc agcgcctcgt 1740
ggccgacggc ctcctcccga agaagggctg cccgcagtcc ggccaggtgg cgatcatcgc 1800
cgacgtggac gagcgcaccc gcaagacggg cgaggccttc gccgccggcc tcgccccgga 1860
ctgcgccatc accgtgcaca cccaggccga cacctcctcc ccggacccgc tcttcaaccc 1920
gctcaagacc ggcgtgtgcc agctcgacgt ggcccaggtg accgacgcca tcctggagcg 1980
cgccggcggc tccatcgccg acttcaccgg ccactaccag accgccttcc gcgagctgga 2040
gcgcgtgctc aacttcccgc agtcgaacct cgccctcaag cgcgagaagc aggacgagtc 2100
cgcctccctc acccaggccc tcccgtccga gctgaaggtg tccgccgaca acgtgtccct 2160
caccggcgcc tggtccctcg cctccatgct caccgaaatc ttcctcctcc agcaggccca 2220
gggcatgccg gagccgggct ggggccgcat caccgactcc caccagtgga acaccctcct 2280
ctccctccac aacgcccagt tcgacctcct ccagcgcacc ccggaggtgg cccgctcccg 2340
cgccaccccg ctcctcgacc tcatcaagac cgccctcacc ccgcacccgc cgcagaagca 2400
ggcctacggc gtgaccctcc cgacctcggt gctcttcatc gccggccacg acaccaacct 2460
cgccaacctc ggcggcgccc tggagctgca gtggaccctc ccgggccagc cggacaacac 2520
cccgccgggc ggcgagctgg tgttcgagcg ctggcgccgc ctctccgaca actcccagtg 2580
gattcaggtg tccctcgtgt tccagaccct ccagcagatg cgcgacaaga ccccgctctt 2640
cctcaacacc ccgccgggcg aggtgaagct caccctggcc ggctgcgagg agcgcaacgc 2700
gcagggcatg tgctccctcg ccggcttcac ccagatcgtg aacgaggccc gcatcccggc 2760
ctgctccctc ggcggcggca gcggcggcgg cagcggcggc ggctgccttt ctttcggaac 2820
tgagatcctt accgttgagt acggaccact tcctattggt aagatcgttt ctgaggaaat 2880
taactgctca gtgtactctg ttgatccaga aggaagagtt tacactcagg ctatcgcaca 2940
atggcacgat aggggtgaac aagaggttct ggagtacgag cttgaagatg gatccgttat 3000
tcgtgctacc tctgaccata gattcttgac tacagattat cagcttctcg ctatcgagga 3060
aatctttgct aggcaacttg atctccttac tttggagaac atcaagcaga cagaagaggc 3120
tcttgacaac cacagacttc cattcccttt gctcgatgct ggaaccatca agtaacctag 3180
gtccccgaat ttccccgatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc 3240
tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat 3300
aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca 3360
attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc 3420
gcgcgcggtg tcatctatgt tactagatcg ggaattg 3457
<210> 133
<211> 3544
<212> DNA
<213> Artificial Sequence
<400> 133
ggtaccaaag taatcatatt attttatgtg tgaatcttct ttactttttc atttgattat 60
gattatgaag gtatgacctt cataaccttc gtccgaaatc cattatatcc aaaggaaaat 120
aatgcttcga aggacgaagg attttgatat ttaacatttt atgttgcctt gttcttaatt 180
catagcattt gagaacaagt ccccaacacc aatctttatc tttactatat taaagcacca 240
gttcaacgat cgtctcgtgt caatattatt aaaaaactcc tacatttctt tataatcaac 300
ccgcactctt ataatctctt ctcttactac tataataaga gagtttatgt acaaaataag 360
gtgaaattat gtataagtgt tctggacctt ggttgttggc tcatattcac acaacctaat 420
caatagaaaa catatgtttt attaaaacaa aatttatcat atatatatat atatatatat 480
atatatatat atatatatat aatataaacc gtagcaatgc acaggcatat gactagtggc 540
aacttaatac catgtgtgta ttaagatgaa taagaggtat ccaaataaat aacttgttcg 600
cttacgtctg gatcgaaagg ggttggaaac gattaaatct cttcctagtc aaaattaaat 660
agaaggagat ttaatcgatt tctcccaatc cccttcgatc caggtgcaac cgaataagtc 720
cttaaatgtt gaggaacacg aaacaaccat gcattggcat gtaaagctcc aagaattcgt 780
tgtatcctta acaactcaca gaacatcaac caaaattgca cgtcaagggt attgggtaag 840
aaacaatcaa acaaatcctc tctgtgtgca aagaaacacg gtgagtcatg ccgagatcat 900
actcatctga tatacatgct tacagctcac aagacattac aaacaactca tattgcatta 960
caaagatcgt ttcatgaaaa ataaaatagg ccggaacagg acaaaaatcc ttgacgtgta 1020
aagtaaattt acaacaaaaa aaaagccata tgtcaagcta aatctaattc gttttacgta 1080
gatcaacaac ctgtagaagg caacaaaact gagccacgca gaagtacaga atgattccag 1140
atgaaccatc gacgtgctac gtaaagagag tgacgagtca tatacatttg gcaagaaacc 1200
atgaagctgc ctacagccgt ctcggtggca taagaacaca agaaattgtg ttaattaatc 1260
aaagctataa ataacgctcg catgcctgtg cacttctcca tcaccaccac tgggtcttca 1320
gaccattagc tttatctact ccagagcgca gaagaacccg atcgacaccg gatccaccat 1380
gagggtgttg ctcgttgccc tcgctctcct ggctctcgct gcgagcgcca ccagcatggt 1440
taaggtgatt ggaagacgtt ctcttggtgt tcaaaggatc ttcgatatcg gattgccaca 1500
agaccacaac tttcttctcg ctaatggtgc catcgctgcc aatagcggcg gcggcagcgg 1560
cggcggcagc accccgcaga gcgccttcgc cgctgcgcag tccgagccgg agctgaagct 1620
ggagtccgtg gtgatcgtgt cgcgccacgg ggtgcgcgcc ccgaccaagt tcacgcagct 1680
catgcaggac gtgaccccgg acgccttcta cacctggccg gtgaagctcg gcgagctgac 1740
cccgcgcggc ggcgagctga tcgcctacct cggccactac tggcgccagc gcctcgtggc 1800
cgacggcctc ctcccgaaga agggctgccc gcagtccggc caggtggcga tcatcgccga 1860
cgtggacgag cgcacccgca agacgggcga ggccttcgcc gccggcctcg ccccggactg 1920
cgccatcacc gtgcacaccc aggccgacac ctcctccccg gacccgctct tcaacccgct 1980
caagaccggc gtgtgccagc tcgacgtggc ccaggtgacc gacgccatcc tggagcgcgc 2040
cggcggctcc atcgccgact tcaccggcca ctaccagacc gccttccgcg agctggagcg 2100
cgtgctcaac ttcccgcagt cgaacctcgc cctcaagcgc gagaagcagg acgagtccgc 2160
ctccctcacc caggccctcc cgtccgagct gaaggtgtcc gccgacaacg tgtccctcac 2220
cggcgcctgg tccctcgcct ccatgctcac cgaaatcttc ctcctccagc aggcccaggg 2280
catgccggag ccgggctggg gccgcatcac cgactcccac cagtggaaca ccctcctctc 2340
cctccacaac gcccagttcg acctcctcca gcgcaccccg gaggtggccc gctcccgcgc 2400
caccccgctc ctcgacctca tcaagaccgc cctcaccccg cacccgccgc agaagcaggc 2460
ctacggcgtg accctcccga cctcggtgct cttcatcgcc ggccacgaca ccaacctcgc 2520
caacctcggc ggcgccctgg agctgcagtg gaccctcccg ggccagccgg acaacacccc 2580
gccgggcggc gagctggtgt tcgagcgctg gcgccgcctc tccgacaact cccagtggat 2640
tcaggtgtcc ctcgtgttcc agaccctcca gcagatgcgc gacaagaccc cgctcttcct 2700
caacaccccg ccgggcgagg tgaagctcac cctggccggc tgcgaggagc gcaacgcgca 2760
gggcatgtgc tccctcgccg gcttcaccca gatcgtgaac gaggcccgca tcccggcctg 2820
ctccctcggc ggcggcagcg gcggcggcag cggcggcggc tgcctttctt tcggaactga 2880
gatccttacc gttgagtacg gaccacttcc tattggtaag atcgtttctg aggaaattaa 2940
ctgctcagtg tactctgttg atccagaagg aagagtttac actcaggcta tcgcacaatg 3000
gcacgatagg ggtgaacaag aggttctgga gtacgagctt gaagatggat ccgttattcg 3060
tgctacctct gaccatagat tcttgactac agattatcag cttctcgcta tcgaggaaat 3120
ctttgctagg caacttgatc tccttacttt ggagaacatc aagcagacag aagaggctct 3180
tgacaaccac agacttccat tccctttgct cgatgctgga accatcaagg acccgaacgg 3240
ctccgagaag gacgagctgt aacctaggtc cccgaatttc cccgatcgtt caaacatttg 3300
gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt 3360
tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag 3420
atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat 3480
atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac tagatcggga 3540
attg 3544
<210> 134
<211> 10
<212> PRT
<213> Artificial Sequence
<400> 134
Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu
1 5 10
<210> 135
<211> 8
<212> PRT
<213> Artificial Sequence
<400> 135
Ser Glu Pro Glu Leu Lys Leu Glu
1 5
<210> 136
<211> 15
<212> PRT
<213> Artificial Sequence
<400> 136
Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val Val Ile Val
1 5 10 15
<210> 137
<211> 1363
<212> DNA
<213> Zea mays
<220>
<221> promoter
<222> (1)..(1363)
<223> Maize gamma zein 27
<400> 137
aaagtaatca tattatttta tgtgtgaatc ttctttactt tttcatttga ttatgattat 60
gaaggtatga ccttcataac cttcgtccga aatccattat atccaaagga aaataatgct 120
tcgaaggacg aaggattttg atatttaaca ttttatgttg ccttgttctt aattcatagc 180
atttgagaac aagtccccaa caccaatctt tatctttact atattaaagc accagttcaa 240
cgatcgtctc gtgtcaatat tattaaaaaa ctcctacatt tctttataat caacccgcac 300
tcttataatc tcttctctta ctactataat aagagagttt atgtacaaaa taaggtgaaa 360
ttatgtataa gtgttctgga ccttggttgt tggctcatat tcacacaacc taatcaatag 420
aaaacatatg ttttattaaa acaaaattta tcatatatat atatatatat atatatatat 480
atatatatat atataatata aaccgtagca atgcacaggc atatgactag tggcaactta 540
ataccatgtg tgtattaaga tgaataagag gtatccaaat aaataacttg ttcgcttacg 600
tctggatcga aaggggttgg aaacgattaa atctcttcct agtcaaaatt aaatagaagg 660
agatttaatc gatttctccc aatccccttc gatccaggtg caaccgaata agtccttaaa 720
tgttgaggaa cacgaaacaa ccatgcattg gcatgtaaag ctccaagaat tcgttgtatc 780
cttaacaact cacagaacat caaccaaaat tgcacgtcaa gggtattggg taagaaacaa 840
tcaaacaaat cctctctgtg tgcaaagaaa cacggtgagt catgccgaga tcatactcat 900
ctgatataca tgcttacagc tcacaagaca ttacaaacaa ctcatattgc attacaaaga 960
tcgtttcatg aaaaataaaa taggccggaa caggacaaaa atccttgacg tgtaaagtaa 1020
atttacaaca aaaaaaaagc catatgtcaa gctaaatcta attcgtttta cgtagatcaa 1080
caacctgtag aaggcaacaa aactgagcca cgcagaagta cagaatgatt ccagatgaac 1140
catcgacgtg ctacgtaaag agagtgacga gtcatataca tttggcaaga aaccatgaag 1200
ctgcctacag ccgtctcggt ggcataagaa cacaagaaat tgtgttaatt aatcaaagct 1260
ataaataacg ctcgcatgcc tgtgcacttc tccatcacca ccactgggtc ttcagaccat 1320
tagctttatc tactccagag cgcagaagaa cccgatcgac acc 1363
<210> 138
<211> 57
<212> DNA
<213> Zea mays
<400> 138
atgcgcgtgc tgctcgtggc cctggccctg ctggctcttg ctgccagcgc cacctct 57
<210> 139
<211> 18
<212> DNA
<213> Artificial Sequence
<400> 139
agcgagaagg acgagctg 18
<210> 140
<211> 6
<212> PRT
<213> Artificial Sequence
<400> 140
Ser Glu Lys Asp Glu Leu
1 5
<210> 141
<211> 276
<212> DNA
<213> Agrobacterium tumefaciens
<400> 141
tccccgaatt tccccgatcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct 60
gttgccggtc ttgcgatgat tatcatataa tttctgttga attacgttaa gcatgtaata 120
attaacatgt aatgcatgac gttatttatg agatgggttt ttatgattag agtcccgcaa 180
ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg 240
cgcgcggtgt catctatgtt actagatcgg gaattg 276
<210> 142
<211> 282
<212> DNA
<213> Artificial Sequence
<400> 142
tgcgtgacag gggacactct catctgcctc gctgacgggc gccgcgttcc tattcaggat 60
ctcgtggggc attcgccgga ggttattgcg gtcgacgata agggccgcct cgtttgcgct 120
aagtcagagg tcatctggaa ggtcggcgag cggtccgttt tcgagatcaa gctggcttcc 180
gggaggagca ttaaggctac cgctgagcac aggctcctgg cgttcaaggg ctggaggcat 240
gttaaggact tcaaagtggg ggataggctc gctattgctc ac 282
<210> 143
<211> 120
<212> DNA
<213> Artificial Sequence
<400> 143
atgtcggacc tgttctggga taggatcgtg tcgattgagg agaaggggtc tgaggaggtc 60
tacgatctca cagttccaaa gtacgcttct tggctcgcgg atggggttgt ttcacataat 120
<210> 144
<211> 288
<212> DNA
<213> Artificial Sequence
<400> 144
tgcctgcacc ctgacacata cgttattctc cctgacgggc gcatgaagaa gatttcggag 60
attgatgagg atgaggttct ctcagtcaac ttcgaggacc tgaagctcta caataagaag 120
atcaagaagt tcaagcacaa ggctccgaag atcctctaca agattaagac cgcgttctcc 180
gagctcatca ccacgggcga gcataagctg ttcgtggtcg agaacgggaa gatcgtcgag 240
aagtgcgtta aggacctcaa tggcagcgag ctgatcgggg ttgtgagg 288
<210> 145
<211> 123
<212> DNA
<213> Artificial Sequence
<400> 145
atggcggata tcgtttggac gaagttcaag attgaggagg tggagagcga tgttgagtat 60
gtgtacgatc tggaggtgga ggactaccac aacttcattg gcaatctcat catcaaccac 120
aac 123
<210> 146
<211> 378
<212> DNA
<213> Artificial Sequence
<400> 146
tgcgttccgc ctgacactct gctcatcctg gagaatgggt tcaagcgcat cgtggacatt 60
aaggtcgggg acaaggtcct gacgcacgag aaccggttca agaaggttga gaaggtgtac 120
aagcgcaggt acatcggcga catcattaag attaaggtgc gctacttccc agaggagatc 180
attctcaccc cagagcaccc tgtctacgct atcaagacgg agaagaggtg cgatggctct 240
catgggatct gcaagttcaa ctgcctcaca cagtacacta atccttcatg caagaagcgg 300
taccgcaagt acaagaggga gtggatcatt gccaaggacc tgaaggtcgg cgatgtgatc 360
gtctacccga ttcccaac 378
<210> 147
<211> 123
<212> DNA
<213> Artificial Sequence
<400> 147
atggggaatt acctgtacgc tcccatcatt aggatcggcc gggagtacta cgacgggttc 60
gtctacaatc tggaggtgga ggatgactct tcatacgtta cagtctcagg cactctgcac 120
aac 123
<210> 148
<211> 330
<212> DNA
<213> Artificial Sequence
<400> 148
tgcctgatgc cgcatgagaa ggtgctgacg gagtacgggg agattaagat tgaggacctg 60
ttcaagatcg ggaaggagat cgtggagaag gacgagctca aggagatcag gaagctgaat 120
attaaggtgc acactctcaa cgagaatggc gagatcaaga tcattaacgc cccatacgtg 180
tggaagctca agcataaggg gaagatgatc aaggtcaagc tgaagaactg gcactcgatc 240
accacgacac cggagcatcc cttcctgacc aacaatggct ggatcaaggc ggagaatatt 300
aagaagggga tgtatgtggc tatccctcgc 330
<210> 149
<211> 117
<212> DNA
<213> Artificial Sequence
<400> 149
atgaacattg cgttcgtcga ggttgaggat gtcgagatca ttgactacga tggctacgtt 60
tacgatctca caacagagac tcataacttc attgctaatg gcatcgtggt tcataat 117
<210> 150
<211> 306
<212> DNA
<213> Artificial Sequence
<400> 150
tgccatccaa aggggacaaa ggtcgtggtc aagggcaagg gcatcgtgaa tattgaggac 60
gttaaggagg ggaattacgt tctcggcatc gacggctggc agaaggttaa gaaggtctgg 120
aagtacgagt acgagggcga gctcattaac gttaatgggc tgaagtgcac accgaaccac 180
aagatccccc tccgctacaa gattaagcat aagaagatca acaagaacga ttacctggtg 240
agggacatct acgcgaagtc gctcctgacc aagttcaagg gcgaggggaa gctcatcctg 300
tgcaag 306
<210> 151
<211> 123
<212> DNA
<213> Artificial Sequence
<400> 151
atgtacgggt tctacgacct cgacgatgtg tgcgtctcac tggagtccta caagggcgag 60
gtgtacgatc tcactctgga gggcaggcct tactacttcg ccaatggcat cctcactcat 120
aat 123
<210> 152
<211> 366
<212> DNA
<213> Artificial Sequence
<400> 152
tgcgtggatt acgagactga ggtcgtgctg gggaatgggg agcggaagaa gatcggggag 60
atcgtggagc gggctattga ggaggctgag aagaacggca agctcgggcg ggttgacgat 120
ggcttctacg ctccgatcga cattgaggtc tactcgctcg atctggagac cctcaaggtt 180
cggaaggcgc gggcaaatat cgcgtggaag cgcacagctc caaagaagat gatgctggtg 240
aagactaggg gcgggaagcg cattagggtc accccgacgc accccttctt cgttctggag 300
gagggcaagg tggctatgag gaaggcccgg gacctggagg agggcaacaa gatcgccacg 360
attgag 366
<210> 153
<211> 123
<212> DNA
<213> Artificial Sequence
<400> 153
atgtccgtga gctgggacga ggtcgcggag atcctggagt acgagccaaa ggatccttgg 60
gtctacgatc tgcaggttcc aggctaccac aacttcctcg ctaatggcat cttcgttcat 120
aat 123
<210> 154
<211> 333
<212> DNA
<213> Artificial Sequence
<400> 154
tgcctcctcc ctgatgagaa ggtcgtggtt ccctcggtcg ggttcgtgac actcaaggag 60
ctgttcgaga cggcttccaa ggtcgtggag cgcgacgatg agaaggagat cagggagctc 120
gacgagcgga ttaccagcgt taacggcgat gggaagacgg gcctggtcaa ggcctcctac 180
gtgtggaagg ttaggcacaa gggcaaggtc atccgggtca agctcaagaa ttggcacggc 240
gttacagtga ctccggagca tcccttcctc accacgaagg ggtggaagag ggctgaccag 300
ctgaggccag gcgattacgt cgcggttcct agg 333
<210> 155
<211> 114
<212> DNA
<213> Artificial Sequence
<400> 155
acgctggtgt tcatccccgt tgagaatgtg gaggaggagg agtacgacgg ctacgtttac 60
gatctcacta cggagactca taacttcatt gctaatggca tcctcgttca taat 114
<210> 156
<211> 363
<212> DNA
<213> Artificial Sequence
<400> 156
tgcgtggacg gggacactct cgtgctgaca aaggagttcg ggctcatcaa gatcaaggac 60
ctctacaaga ttctggacgg caaggggaag aagacagtga acggcaatga ggagtggaca 120
gagctggaga ggccaatcac tctgtacggc tacaaggacg ggaagatcgt cgagattaag 180
gctacccacg tttacaaggg cttctccgcc gggatgatcg agattcggac ccgcacgggc 240
cgcaagatta aggtcacgcc catccataag ctcttcacag gcagggttac taagaatggg 300
ctggagatcc gggaggtcat ggccaaggac ctcaagaagg gcgatcggat cattgtggcg 360
aag 363
<210> 157
<211> 126
<212> DNA
<213> Artificial Sequence
<400> 157
atgacccatg ttctgttcga cgagatcgtg gagattcggt acatctccga gggccaggag 60
gtgtacgacg ttactacgga gactcataat ttcattgggg gcaacatgcc tactctgctc 120
cacaac 126
<210> 158
<211> 333
<212> DNA
<213> Artificial Sequence
<400> 158
tgcctgctgc cggaggagcg ggttattctg cctgactacg ggcctattac tctggaggag 60
ctcttcaata tgacaaagga gacagtgttc aaggacgagg agaaggaggt ccggaagctc 120
ggcatccgca tgccagtggc tggcgtcgat gggcgggtgc gcctgctgga gggcccctac 180
gtttggaagg tgcgctacaa ggggaagatg ctcagggtca agctgaagga ctggcacagc 240
gtggctgtca caccagagca tcccttcctc accacgcggg gctgggtgcg cgctgaccag 300
ctgaagcccg gggattacgt tgccgtgcca aag 333
<210> 159
<211> 117
<212> DNA
<213> Artificial Sequence
<400> 159
atgaacttcg ttttcctgcc ggtggagaag atcgaggagt tcgagtacga tggctacgtc 60
tacgacgtta ctacagagac tcataatttc attgctaatg gcatcctcgt tcataat 117
<210> 160
<211> 363
<212> DNA
<213> Artificial Sequence
<400> 160
tgcgtggacg gggacacact ggtgctgaca aaggagttcg ggctcatcaa gatcaaggag 60
ctctacgaga agctggacgg caaggggcgc aagattgtgg agggcaacga ggagtggacc 120
gagctggaga agccaatcac ggtctacggc tacaaggacg ggaagatcgt tgagattaag 180
gccacccacg tttacaaggg cgtgtccagc gggatggtcg agatcaggac ccggacgggc 240
cggaagatca aggtgacgcc gattcaccgc ctgttcacag gcagggtcac taaggacggg 300
ctgatcctca aggaggtcat ggctatgcat gttaagcccg gcgataggat cgccgtggtc 360
aag 363
<210> 161
<211> 126
<212> DNA
<213> Artificial Sequence
<400> 161
atgcagcata tcattttcga cgaggtcatc gatgtcaggt acattccgga gccccaggag 60
gtgtacgatg ttactacaga gactcataat ttcgtggggg gcaacatgcc aactctgctc 120
cacaat 126
<210> 162
<211> 288
<212> DNA
<213> Artificial Sequence
<400> 162
tgcctcgcgg gggacactct cattacactg gctgacgggc ggcgggttcc tattcgggag 60
ctggtctcgc agcagaattt ctcggtctgg gcgctgaacc cgcagacgta caggctggag 120
agggctcggg tctcccgggc cttctgcaca ggcatcaagc ccgtttacag gctgaccacg 180
aggctcggga ggagcattag ggctactgct aatcaccgct tcctgacccc acagggctgg 240
aagagggtgg acgagctcca gcctggggat tacctggctc tcccaagg 288
<210> 163
<211> 120
<212> DNA
<213> Artificial Sequence
<400> 163
atgtcagacg tctactggga tccgatcgtt tccattgagc ccgacggcgt tgaggaggtg 60
ttcgatctca ctgttccagg gccacataac ttcgttgcta atgacatcat tgctcataat 120
<210> 164
<211> 267
<212> DNA
<213> Artificial Sequence
<400> 164
tgcctcggga aggggacacc ggttatgatg tacgatgggc ggacaaagcc agtggagaag 60
gtggaggtcg gggacaggct catgggggac gatggcagcc caaggacggt gcagtcgctg 120
gccaggggga gggagcagat gtactgggtc cgccagaaga ggggcatgga ctacagggtt 180
aacgagagcc acatcctctc gctgaagaag tctaggaggg agggcgcccg cgacaggggg 240
tcaatcgcgg atatttccgt ccgcgac 267
<210> 165
<211> 114
<212> DNA
<213> Artificial Sequence
<400> 165
atgtggcgga tgaccggcat cgatgtcgag cccgacggcg ttggggatta cttcggcttc 60
actctggatg gcaatgggcg cttcctcctc ggggatggca ctgttactca taat 114
<210> 166
<211> 285
<212> DNA
<213> Artificial Sequence
<400> 166
tgccatcctg cggacactaa ggtcatcgtg aagggcaagg gcatcgttaa tatctcggac 60
gtgaaggagg gggactacat tctcggcatc gacggctggc agcgggtcaa gaaggtttgg 120
aagtaccact acgagggcaa gctcatcaac attaatgggc tgaagtgcac gccgaaccat 180
aaggttcccg tggtcacaga gaatgacagg cagactcgca tcagggattc cctcgccaag 240
agcttcctgt cgggcaaggt caaggggaag atcattacca cgaag 285
<210> 167
<211> 86
<212> DNA
<213> Artificial Sequence
<400> 167
catccgagta ctacaagggc gaggtctacg atctcactct ggagggcaat ccttactact 60
tcgccaatgg catcctcaca cataat 86
<210> 168
<211> 321
<212> DNA
<213> Artificial Sequence
<400> 168
tgcctggaca agacggctct gcggattttc aatcaggggc tgctctacgc ggatgaggtc 60
gtgacaccgg gctcggggga gacagtcggc ctcgggctga cggtcaggaa cggcatcggg 120
gcgtccacag ccattgcgaa tcagccgatg gagctggttg agatcaagct cgctaacggc 180
cggaagctgc gcatgacccc taatcaccgg atgtccgtga agggcaagtg gattcatgcc 240
tgcaacctca agccggggat gctcctggac tacagcatcg gcgagtacca gaagcgcgag 300
gacaccctcc tgattcctct c 321
<210> 169
<211> 147
<212> DNA
<213> Artificial Sequence
<400> 169
atgtcgaagt gcgtcctcaa ctactcgccc tacaagatcg agtctgttaa tattggcgct 60
gtgtgcgact acagctacga tttcgccatc gagggcatca atgataatga ctcttggtac 120
tggcaggggg ctctcaagtc tcacaac 147
<210> 170
<211> 333
<212> DNA
<213> Artificial Sequence
<400> 170
tgcctgctgc cggatgagaa ggttattctc cctgagcatg ggcctattac actcaagggg 60
ctcttcgatc tcgctaagga gacagtcgtg gctgacaacg agaaggagat ccgcaagctg 120
ggcgccaagc tcaccattgt gggcgaggat gggaggctca gggtcctgga gagcccatac 180
gtttggaagg tgcggcaccg cggcaagatg ctgagggtca agctcaagaa ctggcactca 240
gtgtccgtca cgccagagca tcccttcctg accacgcggg gctgggtgcg cgctgaccag 300
ctcaagccgg gggattacgt tgcggtgccc agg 333
<210> 171
<211> 117
<212> DNA
<213> Artificial Sequence
<400> 171
atgaatctcg tcttcatccc ggttgaggac attgaggagt tcgagtacga gggctacgtt 60
tacgacgtta ctacagagac tcataatttc gttgctaatg gcatcctcgt tcataat 117
<210> 172
<211> 297
<212> DNA
<213> Artificial Sequence
<400> 172
tgcctgcctg cgcgggctag ggtcgtggat tggtgcacag ggcgggtcgt tcgggtcggg 60
gagatcgtta ggggggaggc taagggcgtc tgggtggtct ccctggacga ggctaggctg 120
aggctcgttc caaggcctgt tgtggctgct ttcccaagcg gcaaggctca ggtgtacgct 180
ctgaggaccg ctacgggcag ggtgctggag gcgacagcta accacccagt ctacactcca 240
gagggctgga ggccactggg gaccctcgct cctggcgact acgtcgctct gccaagg 297
<210> 173
<211> 120
<212> DNA
<213> Artificial Sequence
<400> 173
atggctgagg tttactggga tcgcgtcgag gcggttgagc cgctcggcga ggaggaggtc 60
ttcgatctca ctgtggaggg cactcatact ttcgttgcgg aggatgttat cgttcataat 120
<210> 174
<211> 42
<212> DNA
<213> Artificial Sequence
<400> 174
atggcccaca tcgtgatggt ggacgcctac aagccgacga ag 42
<210> 175
<211> 39
<212> DNA
<213> Artificial Sequence
<400> 175
gcccacatcg tgatggtgga cgcctacaag ccgacgaag 39
<210> 176
<211> 351
<212> DNA
<213> Artificial Sequence
<400> 176
atgggcgcta tggttgatac cttatcaggt ttatcaagtg agcaaggtca gtccggtgat 60
atgacaattg aagaagatag tgctacccat attaaattct caaaacgtga tgaggacggc 120
aaagagttag ctggtgcaac tatggagttg cgtgattcat ctggtaaaac tattagtaca 180
tggatttcag atggacaagt gaaagatttc tacctgtatc caggaaaata tacatttgtc 240
gaaaccgcag caccagacgg ttatgaggta gcaactgcta ttacctttac agttaatgag 300
caaggtcagg ttactgtaaa tggcaaagca actaaaggtg acgctcatat t 351
<210> 177
<211> 348
<212> DNA
<213> Artificial Sequence
<400> 177
ggcgctatgg ttgatacctt atcaggttta tcaagtgagc aaggtcagtc cggtgatatg 60
acaattgaag aagatagtgc tacccatatt aaattctcaa aacgtgatga ggacggcaaa 120
gagttagctg gtgcaactat ggagttgcgt gattcatctg gtaaaactat tagtacatgg 180
atttcagatg gacaagtgaa agatttctac ctgtatccag gaaaatatac atttgtcgaa 240
accgcagcac cagacggtta tgaggtagca actgctatta cctttacagt taatgagcaa 300
ggtcaggtta ctgtaaatgg caaagcaact aaaggtgacg ctcatatt 348
<210> 178
<211> 102
<212> DNA
<213> Artificial Sequence
<400> 178
atgagggcca agcagctgga ggacaagatt gaggagctgc tgagcaagat ctaccacctg 60
gagaacgaga tagcccgcct gaagaagctg attggcgagc gc 102
<210> 179
<211> 101
<212> DNA
<213> Artificial Sequence
<400> 179
cagctggagg acaagattga ggagctgctg agcaagatct accacctgga gaacgagata 60
gcgaggctga agaagctgat tggcgagcgc accaccccta a 101
<210> 180
<211> 102
<212> DNA
<213> Artificial Sequence
<400> 180
atgagggcca agcagctgga ggacaaggtc gaggagctgc tgagcaagaa ctaccacctg 60
gagaacgagg tcgcccgcct gaagaagctg gtgggcaccc gc 102
<210> 181
<211> 87
<212> DNA
<213> Artificial Sequence
<400> 181
cagctggagg acaaggtcga ggagctgctg agcaagaact accacctgga gaacgaggtc 60
gcgaggctga agaagctggt cggctaa 87
<210> 182
<211> 99
<212> DNA
<213> Artificial Sequence
<400> 182
agcgccgccg aggccgccgc caaggaggcc gccgccaagg aggccgccgc caaggaggcc 60
gccgccaagg ccgttaacac cccgcagagc gccttcgcc 99
<210> 183
<211> 18
<212> DNA
<213> Artificial Sequence
<400> 183
ggttcagggg gttccggt 18
<210> 184
<211> 18
<212> DNA
<213> Artificial Sequence
<400> 184
ggttcagggg gttccggt 18
<210> 185
<211> 1254
<212> DNA
<213> Artificial Sequence
<400> 185
gcgcagtccg agccggagct gaagctggag tccgtggtga tcgtgtcgcg ccacggggtg 60
cgcgccccga ccaaggccac gcagctcatg caggacgtga ccccggacgc ctggccgacc 120
tggccggtga agctcggcga gctgaccccg cgcggcggcg agctgatcgc ctacctcggc 180
cactactggc gccagcgcct cgtggccgac ggcctcctcc cgaagtgcgg ctgcccgcag 240
tccggccagg tggcgatcat cgccgacgtg gacgagcgca cccgcaagac gggcgaggcc 300
ttcgccgccg gcctcgcccc ggactgcgcc atcaccgtgc acacccaggc cgacacctcc 360
tccccggacc cgctcttcaa cccgctcaag accggcgtgt gccagctcga caacgccaac 420
gtgaccgacg ccatcctgga gcgcgccggc ggctccatcg ccgacttcac cggccactac 480
cagaccgcct tccgcgagct ggagcgcgtg ctcaacttcc cgcagtcgaa cctctgcctc 540
aagcgcgaga agcaggacga gtcctgctcc ctcacccagg ccctcccgtc cgagctgaag 600
gtgtccgccg actgcgtgtc cctcaccggc gccgtgtccc tcgcctccat gctcaccgaa 660
atcttcctcc tccagcaggc ccagggcatg ccggagccgg gctggggccg catcaccgac 720
tcccaccagt ggaacaccct cctctccctc cacaacgccc agttcgacct cctccagcgc 780
accccggagg tggcccgctc ccgcgccacc ccgctcctcg acctcatcaa gaccgccctc 840
accccgcacc cgccgcagaa gcaggcctac ggcgtgaccc tcccgacctc ggtgctcttc 900
atcgccggcc acgacaccaa cctcgccaac ctcggcggcg ccctggagct gaactggacc 960
ctcccgggcc agccggacaa caccccgccg ggcggcgagc tggtgttcga gcgctggcgc 1020
cgcctctccg acaactccca gtggattcag gtgtccctcg tgttccagac cctccagcag 1080
atgcgcgaca agaccccgct ctccctcaac accccgccgg gcgaggtgaa gctcaccctg 1140
gccggctgcg aggagcgcaa cgcgcagggc atgtgctccc tcgccggctt cacccagatc 1200
gtgaacgagg cccgcatccc ggcctgctcc ctctccgaga aggacgagct gtaa 1254
<210> 186
<211> 264
<212> DNA
<213> Artificial Sequence
<400> 186
tgtctggacc tgaaaacgca agtgcaaacc ccgcaaggca tgaaggaaat ctcaaacatc 60
caagtcggtg acctggtgct gtcgaatacc ggctataacg aagtgctgaa tgtttttccg 120
aagagcaaaa agaaatctta caagatcacg ctggaagatg gcaaggaaat tatttgcagc 180
gaagaacatc tgttcccgac ccagacgggc gaaatgaata tctccggcgg tctgaaagaa 240
ggcatgtgtc tgtacgtcaa ggaa 264
<210> 187
<211> 88
<212> PRT
<213> Artificial Sequence
<400> 187
Cys Leu Asp Leu Lys Thr Gln Val Gln Thr Pro Gln Gly Met Lys Glu
1 5 10 15
Ile Ser Asn Ile Gln Val Gly Asp Leu Val Leu Ser Asn Thr Gly Tyr
20 25 30
Asn Glu Val Leu Asn Val Phe Pro Lys Ser Lys Lys Lys Ser Tyr Lys
35 40 45
Ile Thr Leu Glu Asp Gly Lys Glu Ile Ile Cys Ser Glu Glu His Leu
50 55 60
Phe Pro Thr Gln Thr Gly Glu Met Asn Ile Ser Gly Gly Leu Lys Glu
65 70 75 80
Gly Met Cys Leu Tyr Val Lys Glu
85
<210> 188
<211> 111
<212> DNA
<213> Artificial Sequence
<400> 188
atgatgctga agaaaattct gaagatcgaa gaactggatg aacgtgaact gattgacatc 60
gaagttagcg gcaaccatct gttttacgcg aatgacattc tgacccacaa c 111
<210> 189
<211> 37
<212> PRT
<213> Artificial Sequence
<400> 189
Met Met Leu Lys Lys Ile Leu Lys Ile Glu Glu Leu Asp Glu Arg Glu
1 5 10 15
Leu Ile Asp Ile Glu Val Ser Gly Asn His Leu Phe Tyr Ala Asn Asp
20 25 30
Ile Leu Thr His Asn
35
<210> 190
<211> 108
<212> DNA
<213> Artificial Sequence
<400> 190
atgacgaaga aaattacgaa gatcgaagaa ctggatgaac gtgaactgat tgacatcgaa 60
gttagcggca accatctgtt ttacgcgaat gacattggga cccacaac 108
<210> 191
<211> 36
<212> PRT
<213> Artificial Sequence
<400> 191
Met Thr Lys Lys Ile Thr Lys Ile Glu Glu Leu Asp Glu Arg Glu Leu
1 5 10 15
Ile Asp Ile Glu Val Ser Gly Asn His Leu Phe Tyr Ala Asn Asp Ile
20 25 30
Gly Thr His Asn
35
<210> 192
<211> 369
<212> DNA
<213> Artificial Sequence
<400> 192
tgcctttctt tcggaactga gatccttacc gttgagtacg gaccacttcc tattggtaag 60
atcgtttctg aggaaattaa ctgctcagtg tactctgttg atccagaagg aagagtttac 120
actcaggcta tcgcacaatg gcacgatagg ggtgaacaag aggttcttga gtacgagctt 180
gaagatggat ccgttattcg tgctacctct gaccatagat tcttgactac agattatcag 240
cttctcgcta tcgaggaaat ctttgctagg caacttgatc tccttacttt ggagaacatc 300
aagcagacag aagaggctct tgacaaccac agacttccat tccctttgct cgatgctgga 360
accatcaag 369
<210> 193
<211> 123
<212> PRT
<213> Artificial Sequence
<400> 193
Cys Leu Ser Phe Gly Thr Glu Ile Leu Thr Val Glu Tyr Gly Pro Leu
1 5 10 15
Pro Ile Gly Lys Ile Val Ser Glu Glu Ile Asn Cys Ser Val Tyr Ser
20 25 30
Val Asp Pro Glu Gly Arg Val Tyr Thr Gln Ala Ile Ala Gln Trp His
35 40 45
Asp Arg Gly Glu Gln Glu Val Leu Glu Tyr Glu Leu Glu Asp Gly Ser
50 55 60
Val Ile Arg Ala Thr Ser Asp His Arg Phe Leu Thr Thr Asp Tyr Gln
65 70 75 80
Leu Leu Ala Ile Glu Glu Ile Phe Ala Arg Gln Leu Asp Leu Leu Thr
85 90 95
Leu Glu Asn Ile Lys Gln Thr Glu Glu Ala Leu Asp Asn His Arg Leu
100 105 110
Pro Phe Pro Leu Leu Asp Ala Gly Thr Ile Lys
115 120
<210> 194
<211> 108
<212> DNA
<213> Artificial Sequence
<400> 194
atggttaagg tgattggaag acgttctctt ggtgttcaaa ggatcttcga tatcggattg 60
ccacaagacc acaactttct tctcgctaat ggtgccatcg ctgccaat 108
<210> 195
<211> 36
<212> PRT
<213> Artificial Sequence
<400> 195
Met Val Lys Val Ile Gly Arg Arg Ser Leu Gly Val Gln Arg Ile Phe
1 5 10 15
Asp Ile Gly Leu Pro Gln Asp His Asn Phe Leu Leu Ala Asn Gly Ala
20 25 30
Ile Ala Ala Asn
35
<210> 196
<211> 255
<212> DNA
<213> Artificial Sequence
<400> 196
atgctgattg aagtgtttag cagtccgatc tgtccgcact gcccaggcgc cgagcgtgtt 60
gtcgaagagg tcgtcgataa actgagctgc gatgatattg aagtgcgcca cattgatgtg 120
acagaagatc cgggcagtgc agagaagtac tctatcatgg cagtgcccac cattgtggta 180
gatggtgagg tggcatttgt tggcgccccg acgacgcagc aatttgagga atatctgcgt 240
aaaaagctta atcgg 255
<210> 197
<211> 85
<212> PRT
<213> Artificial Sequence
<400> 197
Met Leu Ile Glu Val Phe Ser Ser Pro Ile Cys Pro His Cys Pro Gly
1 5 10 15
Ala Glu Arg Val Val Glu Glu Val Val Asp Lys Leu Ser Cys Asp Asp
20 25 30
Ile Glu Val Arg His Ile Asp Val Thr Glu Asp Pro Gly Ser Ala Glu
35 40 45
Lys Tyr Ser Ile Met Ala Val Pro Thr Ile Val Val Asp Gly Glu Val
50 55 60
Ala Phe Val Gly Ala Pro Thr Thr Gln Gln Phe Glu Glu Tyr Leu Arg
65 70 75 80
Lys Lys Leu Asn Arg
85
<210> 198
<211> 12
<212> DNA
<213> Artificial Sequence
<400> 198
gatcctaatg gt 12
<210> 199
<211> 4
<212> PRT
<213> Artificial Sequence
<400> 199
Asp Pro Asn Gly
1
<210> 200
<211> 1752
<212> DNA
<213> Artificial Sequence
<400> 200
atgatgctga agaaaattct gaagatcgaa gaactggatg aacgtgaact gattgacatc 60
gaagttagcg gcaaccatct gttttacgcg aatgacattc tgacccacaa cagcgcagcc 120
gaagccgctg cgaaggaggc agctgcgaaa gaagcggctg caaaagaagc ggcagctaag 180
gctttgaata ccccgcaatc ggctttcgct caatcggaac cggaactgaa actggaaagt 240
gtggttattg tgtctcgtca tggcgttcgc gctccgacca aatttacgca gctgatgcaa 300
gatgtcaccc cggacgcctt ctatacgtgg ccggtgaagc tgggtgaact gaccccgcgt 360
ggcggtgaac tgatcgccta tctgggtcac tactggcgtc agcgcctggt ggcagatggt 420
ctgctgccga aaaagggctg cccgcagagc ggtcaagttg caattatcgc tgatgtcgac 480
gaacgtaccc gcaaaacggg tgaagcattt gcggccggtc tggcaccgga ttgcgccatt 540
accgttcata cgcaggcaga taccagctct ccggacccgc tgttcaaccc gctgaaaacc 600
ggcgtctgtc agctggatgt cgcgcaagtg acggacgcca ttctggaacg tgcaggcggt 660
tccatcgctg attttaccgg tcactaccag acggcattcc gtgaactgga acgcgttctg 720
aactttccgc agtcaaatct ggcgctgaaa cgcgaaaagc aggatgaaag tgcgtccctg 780
acccaagccc tgccgagtga actgaaagtc tccgccgaca atgtgtcact gaccggcgca 840
tggtcactgg cttcgatgct gacggaaatt tttctgctgc agcaagcaca gggtatgccg 900
gaaccgggtt ggggtcgtat caccgattcg catcagtgga acacgctgct gagcctgcac 960
aatgcgcagt tcgacctgct gcaacgtacc ccggaagtgg cacgttcgcg cgccacgccg 1020
ctgctggatc tgattaaaac cgctctgacg ccgcatccgc cgcagaagca agcgtatggc 1080
gtgaccctgc cgacgagcgt tctgtttatc gcgggtcacg acaccaacct ggcaaatctg 1140
ggcggtgctc tggaactgca gtggaccctg ccgggtcaac cggataacac gccgccgggc 1200
ggtgaactgg ttttcgaacg ttggcgtcgc ctgagcgaca attctcagtg gatccaagtt 1260
agcctggtct ttcagaccct gcagcaaatg cgcgataaaa ccccgctgtt cctgaacacg 1320
ccgccgggcg aagtgaagct gaccctggcg ggttgcgaag aacgtaacgc ccagggcatg 1380
tgttctctgg caggttttac ccagattgtt aatgaagcac gcatcccggc ttgtagtctg 1440
gggggcgcag aagcagctgc caaagaggcg gccgcaaagg tcaatctgtg tctggacctg 1500
aaaacgcaag tgcaaacccc gcaaggcatg aaggaaatct caaacatcca agtcggtgac 1560
ctggtgctgt cgaataccgg ctataacgaa gtgctgaatg tttttccgaa gagcaaaaag 1620
aaatcttaca agatcacgct ggaagatggc aaggaaatta tttgcagcga agaacatctg 1680
ttcccgaccc agacgggcga aatgaatatc tccggcggtc tgaaagaagg catgtgtctg 1740
tacgtcaagg aa 1752
<210> 201
<211> 583
<212> PRT
<213> Artificial Sequence
<400> 201
Met Leu Lys Lys Ile Leu Lys Ile Glu Glu Leu Asp Glu Arg Glu Leu
1 5 10 15
Ile Asp Ile Glu Val Ser Gly Asn His Leu Phe Tyr Ala Asn Asp Ile
20 25 30
Leu Thr His Asn Ser Ala Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala
35 40 45
Lys Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Ala Leu Asn Thr Pro
50 55 60
Gln Ser Ala Phe Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val
65 70 75 80
Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe Thr Gln
85 90 95
Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro Val Lys
100 105 110
Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu Gly
115 120 125
His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys Lys
130 135 140
Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp Glu
145 150 155 160
Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro Asp
165 170 175
Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro Asp Pro
180 185 190
Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val Ala Gln
195 200 205
Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp Phe
210 215 220
Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu Asn
225 230 235 240
Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp Glu Ser
245 250 255
Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala Asp
260 265 270
Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu Thr Glu
275 280 285
Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp Gly
290 295 300
Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu His Asn
305 310 315 320
Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser Arg
325 330 335
Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His Pro
340 345 350
Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu Phe
355 360 365
Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu Glu
370 375 380
Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly
385 390 395 400
Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln Trp
405 410 415
Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp Lys
420 425 430
Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr Leu
435 440 445
Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala Gly
450 455 460
Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu Gly
465 470 475 480
Gly Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Val Asn Leu Cys
485 490 495
Leu Asp Leu Lys Thr Gln Val Gln Thr Pro Gln Gly Met Lys Glu Ile
500 505 510
Ser Asn Ile Gln Val Gly Asp Leu Val Leu Ser Asn Thr Gly Tyr Asn
515 520 525
Glu Val Leu Asn Val Phe Pro Lys Ser Lys Lys Lys Ser Tyr Lys Ile
530 535 540
Thr Leu Glu Asp Gly Lys Glu Ile Ile Cys Ser Glu Glu His Leu Phe
545 550 555 560
Pro Thr Gln Thr Gly Glu Met Asn Ile Ser Gly Gly Leu Lys Glu Gly
565 570 575
Met Cys Leu Tyr Val Lys Glu
580
<210> 202
<211> 1692
<212> DNA
<213> Artificial Sequence
<400> 202
atgacgaaga aaattacgaa gatcgaagaa ctggatgaac gtgaactgat tgacatcgaa 60
gttagcggca accatctgtt ttacgcgaat gacattggga cccacaacag cgcagccgaa 120
gccgctgcga aggaggcagc tgcgaaagaa gcggctgcaa aagaagcggc agctaaggct 180
ttgaataccc cgcaatcggc tttcgctcaa tcggaaccgg aactgaaact ggaaagtgtg 240
gttattgtgt ctcgtcatgg cgttcgcgct ccgaccaaat ttacgcagct gatgcaagat 300
gtcaccccgg acgccttcta tacgtggccg gtgaagctgg gtgaactgac cccgcgtggc 360
ggtgaactga tcgcctatct gggtcactac tggcgtcagc gcctggtggc agatggtctg 420
ctgccgaaaa agggctgccc gcagagcggt caagttgcaa ttatcgctga tgtcgacgaa 480
cgtacccgca aaacgggtga agcatttgcg gccggtctgg caccggattg cgccattacc 540
gttcatacgc aggcagatac cagctctccg gacccgctgt tcaacccgct gaaaaccggc 600
gtctgtcagc tggatgtcgc gcaagtgacg gacgccattc tggaacgtgc aggcggttcc 660
atcgctgatt ttaccggtca ctaccagacg gcattccgtg aactggaacg cgttctgaac 720
tttccgcagt caaatctggc gctgaaacgc gaaaagcagg atgaaagtgc gtccctgacc 780
caagccctgc cgagtgaact gaaagtctcc gccgacaatg tgtcactgac cggcgcatgg 840
tcactggctt cgatgctgac ggaaattttt ctgctgcagc aagcacaggg tatgccggaa 900
ccgcagttcg acctgctgca acgtaccccg gaagtggcac gttcgcgcgc cacgccgctg 960
ctggatctga ttaaaaccgc tctgacgccg catccgccgc agaagcaagc gtatggcgtg 1020
accctgccga cgagcgttct gtttatcgcg ggtcacgaca ccaacctggc aaatctgggc 1080
ggtgctctgg aactgcagtg gaccctgccg ggtcaaccgg ataacacgcc gccgggcggt 1140
gaactggttt tcgaacgttg gcgtcgcctg agcgacaatt ctcagtggat ccaagttagc 1200
ctggtctttc agaccctgca gcaaatgcgc gataaaaccc cgctgttcct gaacacgccg 1260
ccgggcgaag tgaagctgac cctggcgggt tgcgaagaac gtaacgccca gggcatgtgt 1320
tctctggcag gttttaccca gattgttaat gaagcacgca tcccggcttg tagtctgggg 1380
ggcgcagaag cagctgccaa agaggcggcc gcaaaggtca atctgtgtct ggacctgaaa 1440
acgcaagtgc aaaccccgca aggcatgaag gaaatctcaa acatccaagt cggtgacctg 1500
gtgctgtcga ataccggcta taacgaagtg ctgaatgttt ttccgaagag caaaaagaaa 1560
tcttacaaga tcacgctgga agatggcaag gaaattattt gcagcgaaga acatctgttc 1620
ccgacccaga cgggcgaaat gaatatctcc ggcggtctga aagaaggcat gtgtctgtac 1680
gtcaaggaat aa 1692
<210> 203
<211> 583
<212> PRT
<213> Artificial Sequence
<400> 203
Met Thr Lys Lys Ile Thr Lys Ile Glu Glu Leu Asp Glu Arg Glu Leu
1 5 10 15
Ile Asp Ile Glu Val Ser Gly Asn His Leu Phe Tyr Ala Asn Asp Ile
20 25 30
Gly Thr His Asn Ser Ala Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala
35 40 45
Lys Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Ala Leu Asn Thr Pro
50 55 60
Gln Ser Ala Phe Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val
65 70 75 80
Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe Thr Gln
85 90 95
Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro Val Lys
100 105 110
Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu Gly
115 120 125
His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys Lys
130 135 140
Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp Glu
145 150 155 160
Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro Asp
165 170 175
Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro Asp Pro
180 185 190
Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val Ala Gln
195 200 205
Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp Phe
210 215 220
Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu Asn
225 230 235 240
Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp Glu Ser
245 250 255
Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala Asp
260 265 270
Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu Thr Glu
275 280 285
Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp Gly
290 295 300
Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu His Asn
305 310 315 320
Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser Arg
325 330 335
Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His Pro
340 345 350
Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu Phe
355 360 365
Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu Glu
370 375 380
Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly
385 390 395 400
Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln Trp
405 410 415
Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp Lys
420 425 430
Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr Leu
435 440 445
Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala Gly
450 455 460
Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu Gly
465 470 475 480
Gly Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Val Asn Leu Cys
485 490 495
Leu Asp Leu Lys Thr Gln Val Gln Thr Pro Gln Gly Met Lys Glu Ile
500 505 510
Ser Asn Ile Gln Val Gly Asp Leu Val Leu Ser Asn Thr Gly Tyr Asn
515 520 525
Glu Val Leu Asn Val Phe Pro Lys Ser Lys Lys Lys Ser Tyr Lys Ile
530 535 540
Thr Leu Glu Asp Gly Lys Glu Ile Ile Cys Ser Glu Glu His Leu Phe
545 550 555 560
Pro Thr Gln Thr Gly Glu Met Asn Ile Ser Gly Gly Leu Lys Glu Gly
565 570 575
Met Cys Leu Tyr Val Lys Glu
580
<210> 204
<211> 2019
<212> DNA
<213> Artificial Sequence
<400> 204
atgctgattg aagtgtttag cagtccgatc tgtccgcact gcccaggcgc cgagcgtgtt 60
gtcgaagagg tcgtcgataa actgagctgc gatgatattg aagtgcgcca cattgatgtg 120
acagaagatc cgggcagtgc agagaagtac tctatcatgg cagtgcccac cattgtggta 180
gatggtgagg tggcatttgt tggcgccccg acgacgcagc aatttgagga atatctgcgt 240
aaaaagctta atcgggatcc taatggtatg acgaagaaaa ttacgaagat cgaagaactg 300
gatgaacgtg aactgattga catcgaagtt agcggcaacc atctgtttta cgcgaatgac 360
attgggaccc acaacagcgc agccgaagcc gctgcgaagg aggcagctgc gaaagaagcg 420
gctgcaaaag aagcggcagc taaggctttg aataccccgc aatcggcttt cgctcaatcg 480
gaaccggaac tgaaactgga aagtgtggtt attgtgtctc gtcatggcgt tcgcgctccg 540
accaaattta cgcagctgat gcaagatgtc accccggacg ccttctatac gtggccggtg 600
aagctgggtg aactgacccc gcgtggcggt gaactgatcg cctatctggg tcactactgg 660
cgtcagcgcc tggtggcaga tggtctgctg ccgaaaaagg gctgcccgca gagcggtcaa 720
gttgcaatta tcgctgatgt cgacgaacgt acccgcaaaa cgggtgaagc atttgcggcc 780
ggtctggcac cggattgcgc cattaccgtt catacgcagg cagataccag ctctccggac 840
ccgctgttca acccgctgaa aaccggcgtc tgtcagctgg atgtcgcgca agtgacggac 900
gccattctgg aacgtgcagg cggttccatc gctgatttta ccggtcacta ccagacggca 960
ttccgtgaac tggaacgcgt tctgaacttt ccgcagtcaa atctggcgct gaaacgcgaa 1020
aagcaggatg aaagtgcgtc cctgacccaa gccctgccga gtgaactgaa agtctccgcc 1080
gacaatgtgt cactgaccgg cgcatggtca ctggcttcga tgctgacgga aatttttctg 1140
ctgcagcaag cacagggtat gccggaaccg ggttggggtc gtatcaccga ttcgcatcag 1200
tggaacacgc tgctgagcct gcacaatgcg cagttcgacc tgctgcaacg taccccggaa 1260
gtggcacgtt cgcgcgccac gccgctgctg gatctgatta aaaccgctct gacgccgcat 1320
ccgccgcaga agcaagcgta tggcgtgacc ctgccgacga gcgttctgtt tatcgcgggt 1380
cacgacacca acctggcaaa tctgggcggt gctctggaac tgcagtggac cctgccgggt 1440
caaccggata acacgccgcc gggcggtgaa ctggttttcg aacgttggcg tcgcctgagc 1500
gacaattctc agtggatcca agttagcctg gtctttcaga ccctgcagca aatgcgcgat 1560
aaaaccccgc tgttcctgaa cacgccgccg ggcgaagtga agctgaccct ggcgggttgc 1620
gaagaacgta acgcccaggg catgtgttct ctggcaggtt ttacccagat tgttaatgaa 1680
gcacgcatcc cggcttgtag tctggggggc gcagaagcag ctgccaaaga ggcggccgca 1740
aaggtcaatc tgtgtctgga cctgaaaacg caagtgcaaa ccccgcaagg catgaaggaa 1800
atctcaaaca tccaagtcgg tgacctggtg ctgtcgaata ccggctataa cgaagtgctg 1860
aatgtttttc cgaagagcaa aaagaaatct tacaagatca cgctggaaga tggcaaggaa 1920
attatttgca gcgaagaaca tctgttcccg acccagacgg gcgaaatgaa tatctccggc 1980
ggtctgaaag aaggcatgtg tctgtacgtc aaggaataa 2019
<210> 205
<211> 672
<212> PRT
<213> Artificial Sequence
<400> 205
Met Leu Ile Glu Val Phe Ser Ser Pro Ile Cys Pro His Cys Pro Gly
1 5 10 15
Ala Glu Arg Val Val Glu Glu Val Val Asp Lys Leu Ser Cys Asp Asp
20 25 30
Ile Glu Val Arg His Ile Asp Val Thr Glu Asp Pro Gly Ser Ala Glu
35 40 45
Lys Tyr Ser Ile Met Ala Val Pro Thr Ile Val Val Asp Gly Glu Val
50 55 60
Ala Phe Val Gly Ala Pro Thr Thr Gln Gln Phe Glu Glu Tyr Leu Arg
65 70 75 80
Lys Lys Leu Asn Arg Asp Pro Asn Gly Met Thr Lys Lys Ile Thr Lys
85 90 95
Ile Glu Glu Leu Asp Glu Arg Glu Leu Ile Asp Ile Glu Val Ser Gly
100 105 110
Asn His Leu Phe Tyr Ala Asn Asp Ile Gly Thr His Asn Ser Ala Ala
115 120 125
Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Glu
130 135 140
Ala Ala Ala Lys Ala Leu Asn Thr Pro Gln Ser Ala Phe Ala Gln Ser
145 150 155 160
Glu Pro Glu Leu Lys Leu Glu Ser Val Val Ile Val Ser Arg His Gly
165 170 175
Val Arg Ala Pro Thr Lys Phe Thr Gln Leu Met Gln Asp Val Thr Pro
180 185 190
Asp Ala Phe Tyr Thr Trp Pro Val Lys Leu Gly Glu Leu Thr Pro Arg
195 200 205
Gly Gly Glu Leu Ile Ala Tyr Leu Gly His Tyr Trp Arg Gln Arg Leu
210 215 220
Val Ala Asp Gly Leu Leu Pro Lys Lys Gly Cys Pro Gln Ser Gly Gln
225 230 235 240
Val Ala Ile Ile Ala Asp Val Asp Glu Arg Thr Arg Lys Thr Gly Glu
245 250 255
Ala Phe Ala Ala Gly Leu Ala Pro Asp Cys Ala Ile Thr Val His Thr
260 265 270
Gln Ala Asp Thr Ser Ser Pro Asp Pro Leu Phe Asn Pro Leu Lys Thr
275 280 285
Gly Val Cys Gln Leu Asp Val Ala Gln Val Thr Asp Ala Ile Leu Glu
290 295 300
Arg Ala Gly Gly Ser Ile Ala Asp Phe Thr Gly His Tyr Gln Thr Ala
305 310 315 320
Phe Arg Glu Leu Glu Arg Val Leu Asn Phe Pro Gln Ser Asn Leu Ala
325 330 335
Leu Lys Arg Glu Lys Gln Asp Glu Ser Ala Ser Leu Thr Gln Ala Leu
340 345 350
Pro Ser Glu Leu Lys Val Ser Ala Asp Asn Val Ser Leu Thr Gly Ala
355 360 365
Trp Ser Leu Ala Ser Met Leu Thr Glu Ile Phe Leu Leu Gln Gln Ala
370 375 380
Gln Gly Met Pro Glu Pro Gly Trp Gly Arg Ile Thr Asp Ser His Gln
385 390 395 400
Trp Asn Thr Leu Leu Ser Leu His Asn Ala Gln Phe Asp Leu Leu Gln
405 410 415
Arg Thr Pro Glu Val Ala Arg Ser Arg Ala Thr Pro Leu Leu Asp Leu
420 425 430
Ile Lys Thr Ala Leu Thr Pro His Pro Pro Gln Lys Gln Ala Tyr Gly
435 440 445
Val Thr Leu Pro Thr Ser Val Leu Phe Ile Ala Gly His Asp Thr Asn
450 455 460
Leu Ala Asn Leu Gly Gly Ala Leu Glu Leu Gln Trp Thr Leu Pro Gly
465 470 475 480
Gln Pro Asp Asn Thr Pro Pro Gly Gly Glu Leu Val Phe Glu Arg Trp
485 490 495
Arg Arg Leu Ser Asp Asn Ser Gln Trp Ile Gln Val Ser Leu Val Phe
500 505 510
Gln Thr Leu Gln Gln Met Arg Asp Lys Thr Pro Leu Phe Leu Asn Thr
515 520 525
Pro Pro Gly Glu Val Lys Leu Thr Leu Ala Gly Cys Glu Glu Arg Asn
530 535 540
Ala Gln Gly Met Cys Ser Leu Ala Gly Phe Thr Gln Ile Val Asn Glu
545 550 555 560
Ala Arg Ile Pro Ala Cys Ser Leu Gly Gly Ala Glu Ala Ala Ala Lys
565 570 575
Glu Ala Ala Ala Lys Val Asn Leu Cys Leu Asp Leu Lys Thr Gln Val
580 585 590
Gln Thr Pro Gln Gly Met Lys Glu Ile Ser Asn Ile Gln Val Gly Asp
595 600 605
Leu Val Leu Ser Asn Thr Gly Tyr Asn Glu Val Leu Asn Val Phe Pro
610 615 620
Lys Ser Lys Lys Lys Ser Tyr Lys Ile Thr Leu Glu Asp Gly Lys Glu
625 630 635 640
Ile Ile Cys Ser Glu Glu His Leu Phe Pro Thr Gln Thr Gly Glu Met
645 650 655
Asn Ile Ser Gly Gly Leu Lys Glu Gly Met Cys Leu Tyr Val Lys Glu
660 665 670
<210> 206
<211> 1992
<212> DNA
<213> Artificial Sequence
<400> 206
atgctgattg aagtgtttag cagtccgatc tgtccgcact gcccaggcgc cgagcgtgtt 60
gtcgaagagg tcgtcgataa actgagctgc gatgatattg aagtgcgcca cattgatgtg 120
acagaagatc cgggcagtgc agagaagtac tctatcatgg cagtgcccac cattgtggta 180
gatggtgagg tggcatttgt tggcgccccg acgacgcagc aatttgagga atatctgcgt 240
aaaaagctta atcgggatcc taatggtatg acgaagaaaa ttacgaagat cgaagaactg 300
gatgaacgtg aactgattga catcgaagtt agcggcaacc atctgtttta cgcgaatgac 360
attgggaccc acaacagcgc ctttgcacaa tcggaaccgg aactgaaact ggaaagtgtg 420
gttattgtgt ctcgtcatgg cgttcgcgct ccgaccaaat ttacgcagct gatgcaagat 480
gtcaccccgg acgccttcta tacgtggccg gtgaagctgg gtgaactgac cccgcgtggc 540
ggtgaactga tcgcctatct gggtcactac tggcgtcagc gcctggtggc agatggtctg 600
ctgccgaaaa agggctgccc gcagagcggt caagttgcaa ttatcgctga tgtcgacgaa 660
cgtacccgca aaacgggtga agcatttgcg gccggtctgg caccggattg cgccattacc 720
gttcatacgc aggcagatac cagctctccg gacccgctgt tcaacccgct gaaaaccggc 780
gtctgtcagc tggatgtcgc gcaagtgacg gacgccattc tggaacgtgc aggcggttcc 840
atcgctgatt ttaccggtca ctaccagacg gcattccgtg aactggaacg cgttctgaac 900
tttccgcagt caaatctggc gctgaaacgc gaaaagcagg atgaaagtgc gtccctgacc 960
caagccctgc cgagtgaact gaaagtctcc gccgacaatg tgtcactgac cggcgcatgg 1020
tcactggctt cgatgctgac ggaaattttt ctgctgcagc aagcacaggg tatgccggaa 1080
ccgggttggg gtcgtatcac cgattcgcat cagtggaaca cgctgctgag cctgcacaat 1140
gcgcagttcg acctgctgca acgtaccccg gaagtggcac gttcgcgcgc cacgccgctg 1200
ctggatctga ttaaaaccgc tctgacgccg catccgccgc agaagcaagc gtatggcgtg 1260
accctgccga cgagcgttct gtttatcgcg ggtcacgaca ccaacctggc aaatctgggc 1320
ggtgctctgg aactgcagtg gaccctgccg ggtcaaccgg ataacacgcc gccgggcggt 1380
gaactggttt tcgaacgttg gcgtcgcctg agcgacaatt ctcagtggat ccaagttagc 1440
ctggtctttc agaccctgca gcaaatgcgc gataaaaccc cgctgttcct gaacacgccg 1500
ccgggcgaag tgaagctgac cctggcgggt tgcgaagaac gtaacgccca gggcatgtgt 1560
tctctggcag gttttaccca gattgttaat gaagcacgca tcccggcttg tagtctgggt 1620
gcagctccag cggccgcacc ggctaaacag gaagcggcag ctccggctcc tgcagcgaag 1680
gcggaagcac cggccgcagc tcctgcggca aaagcgaccc cgcagtgtct ggacctgaaa 1740
acgcaagtgc aaaccccgca aggcatgaag gaaatctcaa acatccaagt cggtgacctg 1800
gtgctgtcga ataccggcta taacgaagtg ctgaatgttt ttccgaagag caaaaagaaa 1860
tcttacaaga tcacgctgga agatggcaag gaaattattt gcagcgaaga acatctgttc 1920
ccgacccaga cgggcgaaat gaatatctcc ggcggtctga aagaaggcat gtgtctgtac 1980
gtcaaggaat aa 1992
<210> 207
<211> 663
<212> PRT
<213> Artificial Sequence
<400> 207
Met Leu Ile Glu Val Phe Ser Ser Pro Ile Cys Pro His Cys Pro Gly
1 5 10 15
Ala Glu Arg Val Val Glu Glu Val Val Asp Lys Leu Ser Cys Asp Asp
20 25 30
Ile Glu Val Arg His Ile Asp Val Thr Glu Asp Pro Gly Ser Ala Glu
35 40 45
Lys Tyr Ser Ile Met Ala Val Pro Thr Ile Val Val Asp Gly Glu Val
50 55 60
Ala Phe Val Gly Ala Pro Thr Thr Gln Gln Phe Glu Glu Tyr Leu Arg
65 70 75 80
Lys Lys Leu Asn Arg Asp Pro Asn Gly Met Thr Lys Lys Ile Thr Lys
85 90 95
Ile Glu Glu Leu Asp Glu Arg Glu Leu Ile Asp Ile Glu Val Ser Gly
100 105 110
Asn His Leu Phe Tyr Ala Asn Asp Ile Gly Thr His Asn Ser Ala Phe
115 120 125
Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val Val Ile Val Ser
130 135 140
Arg His Gly Val Arg Ala Pro Thr Lys Phe Thr Gln Leu Met Gln Asp
145 150 155 160
Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro Val Lys Leu Gly Glu Leu
165 170 175
Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu Gly His Tyr Trp Arg
180 185 190
Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys Lys Gly Cys Pro Gln
195 200 205
Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp Glu Arg Thr Arg Lys
210 215 220
Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro Asp Cys Ala Ile Thr
225 230 235 240
Val His Thr Gln Ala Asp Thr Ser Ser Pro Asp Pro Leu Phe Asn Pro
245 250 255
Leu Lys Thr Gly Val Cys Gln Leu Asp Val Ala Gln Val Thr Asp Ala
260 265 270
Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp Phe Thr Gly His Tyr
275 280 285
Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu Asn Phe Pro Gln Ser
290 295 300
Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp Glu Ser Ala Ser Leu Thr
305 310 315 320
Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala Asp Asn Val Ser Leu
325 330 335
Thr Gly Ala Trp Ser Leu Ala Ser Met Leu Thr Glu Ile Phe Leu Leu
340 345 350
Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp Gly Arg Ile Thr Asp
355 360 365
Ser His Gln Trp Asn Thr Leu Leu Ser Leu His Asn Ala Gln Phe Asp
370 375 380
Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser Arg Ala Thr Pro Leu
385 390 395 400
Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His Pro Pro Gln Lys Gln
405 410 415
Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu Phe Ile Ala Gly His
420 425 430
Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu Glu Leu Gln Trp Thr
435 440 445
Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly Glu Leu Val Phe
450 455 460
Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln Trp Ile Gln Val Ser
465 470 475 480
Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp Lys Thr Pro Leu Phe
485 490 495
Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr Leu Ala Gly Cys Glu
500 505 510
Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala Gly Phe Thr Gln Ile
515 520 525
Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu Gly Ala Ala Pro Ala
530 535 540
Ala Ala Pro Ala Lys Gln Glu Ala Ala Ala Pro Ala Pro Ala Ala Lys
545 550 555 560
Ala Glu Ala Pro Ala Ala Ala Pro Ala Ala Lys Ala Thr Pro Gln Cys
565 570 575
Leu Asp Leu Lys Thr Gln Val Gln Thr Pro Gln Gly Met Lys Glu Ile
580 585 590
Ser Asn Ile Gln Val Gly Asp Leu Val Leu Ser Asn Thr Gly Tyr Asn
595 600 605
Glu Val Leu Asn Val Phe Pro Lys Ser Lys Lys Lys Ser Tyr Lys Ile
610 615 620
Thr Leu Glu Asp Gly Lys Glu Ile Ile Cys Ser Glu Glu His Leu Phe
625 630 635 640
Pro Thr Gln Thr Gly Glu Met Asn Ile Ser Gly Gly Leu Lys Glu Gly
645 650 655
Met Cys Leu Tyr Val Lys Glu
660
<210> 208
<211> 1992
<212> DNA
<213> Artificial Sequence
<400> 208
atgctgattg aagtgtttag cagtccgatc tgtccgcact gcccaggcgc cgagcgtgtt 60
gtcgaagagg tcgtcgataa actgagctgc gatgatattg aagtgcgcca cattgatgtg 120
acagaagatc cgggcagtgc agagaagtac tctatcatgg cagtgcccac cattgtggta 180
gatggtgagg tggcatttgt tggcgccccg acgacgcagc aatttgagga atatctgcgt 240
aaaaagctta atcgggatcc taatggtatg acgaagaaaa ttacgaagat cgaagaactg 300
gatgaacgtg aactgattga catcgaagtt agcggcaacc atctgtttta cgcgaatgac 360
attgggaccc acgccagcgc ctttgcacaa tcggaaccgg aactgaaact ggaaagtgtg 420
gttattgtgt ctcgtcatgg cgttcgcgct ccgaccaaat ttacgcagct gatgcaagat 480
gtcaccccgg acgccttcta tacgtggccg gtgaagctgg gtgaactgac cccgcgtggc 540
ggtgaactga tcgcctatct gggtcactac tggcgtcagc gcctggtggc agatggtctg 600
ctgccgaaaa agggctgccc gcagagcggt caagttgcaa ttatcgctga tgtcgacgaa 660
cgtacccgca aaacgggtga agcatttgcg gccggtctgg caccggattg cgccattacc 720
gttcatacgc aggcagatac cagctctccg gacccgctgt tcaacccgct gaaaaccggc 780
gtctgtcagc tggatgtcgc gcaagtgacg gacgccattc tggaacgtgc aggcggttcc 840
atcgctgatt ttaccggtca ctaccagacg gcattccgtg aactggaacg cgttctgaac 900
tttccgcagt caaatctggc gctgaaacgc gaaaagcagg atgaaagtgc gtccctgacc 960
caagccctgc cgagtgaact gaaagtctcc gccgacaatg tgtcactgac cggcgcatgg 1020
tcactggctt cgatgctgac ggaaattttt ctgctgcagc aagcacaggg tatgccggaa 1080
ccgggttggg gtcgtatcac cgattcgcat cagtggaaca cgctgctgag cctgcacaat 1140
gcgcagttcg acctgctgca acgtaccccg gaagtggcac gttcgcgcgc cacgccgctg 1200
ctggatctga ttaaaaccgc tctgacgccg catccgccgc agaagcaagc gtatggcgtg 1260
accctgccga cgagcgttct gtttatcgcg ggtcacgaca ccaacctggc aaatctgggc 1320
ggtgctctgg aactgcagtg gaccctgccg ggtcaaccgg ataacacgcc gccgggcggt 1380
gaactggttt tcgaacgttg gcgtcgcctg agcgacaatt ctcagtggat ccaagttagc 1440
ctggtctttc agaccctgca gcaaatgcgc gataaaaccc cgctgttcct gaacacgccg 1500
ccgggcgaag tgaagctgac cctggcgggt tgcgaagaac gtaacgccca gggcatgtgt 1560
tctctggcag gttttaccca gattgttaat gaagcacgca tcccggcttg tagtctgggt 1620
gcagctccag cggccgcacc ggctaaacag gaagcggcag ctccggctcc tgcagcgaag 1680
gcggaagcac cggccgcagc tcctgcggca aaagcgaccc cgcagtgtct ggacctgaaa 1740
acgcaagtgc aaaccccgca aggcatgaag gaaatctcaa acatccaagt cggtgacctg 1800
gtgctgtcga ataccggcta taacgaagtg ctgaatgttt ttccgaagag caaaaagaaa 1860
tcttacaaga tcacgctgga agatggcaag gaaattattt gcagcgaaga acatctgttc 1920
ccgacccaga cgggcgaaat gaatatctcc ggcggtctga aagaaggcat gtgtctgtac 1980
gtcaaggaat aa 1992
<210> 209
<211> 663
<212> PRT
<213> Artificial Sequence
<400> 209
Met Leu Ile Glu Val Phe Ser Ser Pro Ile Cys Pro His Cys Pro Gly
1 5 10 15
Ala Glu Arg Val Val Glu Glu Val Val Asp Lys Leu Ser Cys Asp Asp
20 25 30
Ile Glu Val Arg His Ile Asp Val Thr Glu Asp Pro Gly Ser Ala Glu
35 40 45
Lys Tyr Ser Ile Met Ala Val Pro Thr Ile Val Val Asp Gly Glu Val
50 55 60
Ala Phe Val Gly Ala Pro Thr Thr Gln Gln Phe Glu Glu Tyr Leu Arg
65 70 75 80
Lys Lys Leu Asn Arg Asp Pro Asn Gly Met Thr Lys Lys Ile Thr Lys
85 90 95
Ile Glu Glu Leu Asp Glu Arg Glu Leu Ile Asp Ile Glu Val Ser Gly
100 105 110
Asn His Leu Phe Tyr Ala Asn Asp Ile Gly Thr His Ala Ser Ala Phe
115 120 125
Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val Val Ile Val Ser
130 135 140
Arg His Gly Val Arg Ala Pro Thr Lys Phe Thr Gln Leu Met Gln Asp
145 150 155 160
Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro Val Lys Leu Gly Glu Leu
165 170 175
Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu Gly His Tyr Trp Arg
180 185 190
Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys Lys Gly Cys Pro Gln
195 200 205
Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp Glu Arg Thr Arg Lys
210 215 220
Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro Asp Cys Ala Ile Thr
225 230 235 240
Val His Thr Gln Ala Asp Thr Ser Ser Pro Asp Pro Leu Phe Asn Pro
245 250 255
Leu Lys Thr Gly Val Cys Gln Leu Asp Val Ala Gln Val Thr Asp Ala
260 265 270
Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp Phe Thr Gly His Tyr
275 280 285
Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu Asn Phe Pro Gln Ser
290 295 300
Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp Glu Ser Ala Ser Leu Thr
305 310 315 320
Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala Asp Asn Val Ser Leu
325 330 335
Thr Gly Ala Trp Ser Leu Ala Ser Met Leu Thr Glu Ile Phe Leu Leu
340 345 350
Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp Gly Arg Ile Thr Asp
355 360 365
Ser His Gln Trp Asn Thr Leu Leu Ser Leu His Asn Ala Gln Phe Asp
370 375 380
Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser Arg Ala Thr Pro Leu
385 390 395 400
Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His Pro Pro Gln Lys Gln
405 410 415
Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu Phe Ile Ala Gly His
420 425 430
Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu Glu Leu Gln Trp Thr
435 440 445
Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly Glu Leu Val Phe
450 455 460
Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln Trp Ile Gln Val Ser
465 470 475 480
Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp Lys Thr Pro Leu Phe
485 490 495
Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr Leu Ala Gly Cys Glu
500 505 510
Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala Gly Phe Thr Gln Ile
515 520 525
Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu Gly Ala Ala Pro Ala
530 535 540
Ala Ala Pro Ala Lys Gln Glu Ala Ala Ala Pro Ala Pro Ala Ala Lys
545 550 555 560
Ala Glu Ala Pro Ala Ala Ala Pro Ala Ala Lys Ala Thr Pro Gln Cys
565 570 575
Leu Asp Leu Lys Thr Gln Val Gln Thr Pro Gln Gly Met Lys Glu Ile
580 585 590
Ser Asn Ile Gln Val Gly Asp Leu Val Leu Ser Asn Thr Gly Tyr Asn
595 600 605
Glu Val Leu Asn Val Phe Pro Lys Ser Lys Lys Lys Ser Tyr Lys Ile
610 615 620
Thr Leu Glu Asp Gly Lys Glu Ile Ile Cys Ser Glu Glu His Leu Phe
625 630 635 640
Pro Thr Gln Thr Gly Glu Met Asn Ile Ser Gly Gly Leu Lys Glu Gly
645 650 655
Met Cys Leu Tyr Val Lys Glu
660
<210> 210
<211> 1992
<212> DNA
<213> Artificial Sequence
<400> 210
atgctgattg aagtgtttag cagtccgatc tgtccgcact gcccaggcgc cgagcgtgtt 60
gtcgaagagg tcgtcgataa actgagctgc gatgatattg aagtgcgcca cattgatgtg 120
acagaagatc cgggcagtgc agagaagtac tctatcatgg cagtgcccac cattgtggta 180
gatggtgagg tggcatttgt tggcgccccg acgacgcagc aatttgagga atatctgcgt 240
aaaaagctta atcgggatcc taatggtatg acgaagaaaa ttacgaagat cgaagaactg 300
gatgaacgtg aactgattga catcgaagtt agcggcaacc atctgtttta cgcgaatgac 360
attgggaccc acaacgccgc ctttgcacaa tcggaaccgg aactgaaact ggaaagtgtg 420
gttattgtgt ctcgtcatgg cgttcgcgct ccgaccaaat ttacgcagct gatgcaagat 480
gtcaccccgg acgccttcta tacgtggccg gtgaagctgg gtgaactgac cccgcgtggc 540
ggtgaactga tcgcctatct gggtcactac tggcgtcagc gcctggtggc agatggtctg 600
ctgccgaaaa agggctgccc gcagagcggt caagttgcaa ttatcgctga tgtcgacgaa 660
cgtacccgca aaacgggtga agcatttgcg gccggtctgg caccggattg cgccattacc 720
gttcatacgc aggcagatac cagctctccg gacccgctgt tcaacccgct gaaaaccggc 780
gtctgtcagc tggatgtcgc gcaagtgacg gacgccattc tggaacgtgc aggcggttcc 840
atcgctgatt ttaccggtca ctaccagacg gcattccgtg aactggaacg cgttctgaac 900
tttccgcagt caaatctggc gctgaaacgc gaaaagcagg atgaaagtgc gtccctgacc 960
caagccctgc cgagtgaact gaaagtctcc gccgacaatg tgtcactgac cggcgcatgg 1020
tcactggctt cgatgctgac ggaaattttt ctgctgcagc aagcacaggg tatgccggaa 1080
ccgggttggg gtcgtatcac cgattcgcat cagtggaaca cgctgctgag cctgcacaat 1140
gcgcagttcg acctgctgca acgtaccccg gaagtggcac gttcgcgcgc cacgccgctg 1200
ctggatctga ttaaaaccgc tctgacgccg catccgccgc agaagcaagc gtatggcgtg 1260
accctgccga cgagcgttct gtttatcgcg ggtcacgaca ccaacctggc aaatctgggc 1320
ggtgctctgg aactgcagtg gaccctgccg ggtcaaccgg ataacacgcc gccgggcggt 1380
gaactggttt tcgaacgttg gcgtcgcctg agcgacaatt ctcagtggat ccaagttagc 1440
ctggtctttc agaccctgca gcaaatgcgc gataaaaccc cgctgttcct gaacacgccg 1500
ccgggcgaag tgaagctgac cctggcgggt tgcgaagaac gtaacgccca gggcatgtgt 1560
tctctggcag gttttaccca gattgttaat gaagcacgca tcccggcttg tagtctgggt 1620
gcagctccag cggccgcacc ggctaaacag gaagcggcag ctccggctcc tgcagcgaag 1680
gcggaagcac cggccgcagc tcctgcggca aaagcgaccc cgcagtgtct ggacctgaaa 1740
acgcaagtgc aaaccccgca aggcatgaag gaaatctcaa acatccaagt cggtgacctg 1800
gtgctgtcga ataccggcta taacgaagtg ctgaatgttt ttccgaagag caaaaagaaa 1860
tcttacaaga tcacgctgga agatggcaag gaaattattt gcagcgaaga acatctgttc 1920
ccgacccaga cgggcgaaat gaatatctcc ggcggtctga aagaaggcat gtgtctgtac 1980
gtcaaggaat aa 1992
<210> 211
<211> 663
<212> PRT
<213> Artificial Sequence
<400> 211
Met Leu Ile Glu Val Phe Ser Ser Pro Ile Cys Pro His Cys Pro Gly
1 5 10 15
Ala Glu Arg Val Val Glu Glu Val Val Asp Lys Leu Ser Cys Asp Asp
20 25 30
Ile Glu Val Arg His Ile Asp Val Thr Glu Asp Pro Gly Ser Ala Glu
35 40 45
Lys Tyr Ser Ile Met Ala Val Pro Thr Ile Val Val Asp Gly Glu Val
50 55 60
Ala Phe Val Gly Ala Pro Thr Thr Gln Gln Phe Glu Glu Tyr Leu Arg
65 70 75 80
Lys Lys Leu Asn Arg Asp Pro Asn Gly Met Thr Lys Lys Ile Thr Lys
85 90 95
Ile Glu Glu Leu Asp Glu Arg Glu Leu Ile Asp Ile Glu Val Ser Gly
100 105 110
Asn His Leu Phe Tyr Ala Asn Asp Ile Gly Thr His Asn Ala Ala Phe
115 120 125
Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val Val Ile Val Ser
130 135 140
Arg His Gly Val Arg Ala Pro Thr Lys Phe Thr Gln Leu Met Gln Asp
145 150 155 160
Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro Val Lys Leu Gly Glu Leu
165 170 175
Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu Gly His Tyr Trp Arg
180 185 190
Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys Lys Gly Cys Pro Gln
195 200 205
Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp Glu Arg Thr Arg Lys
210 215 220
Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro Asp Cys Ala Ile Thr
225 230 235 240
Val His Thr Gln Ala Asp Thr Ser Ser Pro Asp Pro Leu Phe Asn Pro
245 250 255
Leu Lys Thr Gly Val Cys Gln Leu Asp Val Ala Gln Val Thr Asp Ala
260 265 270
Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp Phe Thr Gly His Tyr
275 280 285
Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu Asn Phe Pro Gln Ser
290 295 300
Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp Glu Ser Ala Ser Leu Thr
305 310 315 320
Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala Asp Asn Val Ser Leu
325 330 335
Thr Gly Ala Trp Ser Leu Ala Ser Met Leu Thr Glu Ile Phe Leu Leu
340 345 350
Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp Gly Arg Ile Thr Asp
355 360 365
Ser His Gln Trp Asn Thr Leu Leu Ser Leu His Asn Ala Gln Phe Asp
370 375 380
Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser Arg Ala Thr Pro Leu
385 390 395 400
Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His Pro Pro Gln Lys Gln
405 410 415
Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu Phe Ile Ala Gly His
420 425 430
Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu Glu Leu Gln Trp Thr
435 440 445
Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly Glu Leu Val Phe
450 455 460
Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln Trp Ile Gln Val Ser
465 470 475 480
Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp Lys Thr Pro Leu Phe
485 490 495
Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr Leu Ala Gly Cys Glu
500 505 510
Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala Gly Phe Thr Gln Ile
515 520 525
Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu Gly Ala Ala Pro Ala
530 535 540
Ala Ala Pro Ala Lys Gln Glu Ala Ala Ala Pro Ala Pro Ala Ala Lys
545 550 555 560
Ala Glu Ala Pro Ala Ala Ala Pro Ala Ala Lys Ala Thr Pro Gln Cys
565 570 575
Leu Asp Leu Lys Thr Gln Val Gln Thr Pro Gln Gly Met Lys Glu Ile
580 585 590
Ser Asn Ile Gln Val Gly Asp Leu Val Leu Ser Asn Thr Gly Tyr Asn
595 600 605
Glu Val Leu Asn Val Phe Pro Lys Ser Lys Lys Lys Ser Tyr Lys Ile
610 615 620
Thr Leu Glu Asp Gly Lys Glu Ile Ile Cys Ser Glu Glu His Leu Phe
625 630 635 640
Pro Thr Gln Thr Gly Glu Met Asn Ile Ser Gly Gly Leu Lys Glu Gly
645 650 655
Met Cys Leu Tyr Val Lys Glu
660
<210> 212
<211> 1692
<212> DNA
<213> Artificial Sequence
<400> 212
atgacgaaga aaattacgaa gatcgaagaa ctggatgaac gtgaactgat tgacatcgaa 60
gttagcggca accatctgtt ttacgcgaat gacattggga cccacgccag cgcagccgaa 120
gccgctgcga aggaggcagc tgcgaaagaa gcggctgcaa aagaagcggc agctaaggct 180
ttgaataccc cgcaatcggc tttcgctcaa tcggaaccgg aactgaaact ggaaagtgtg 240
gttattgtgt ctcgtcatgg cgttcgcgct ccgaccaaat ttacgcagct gatgcaagat 300
gtcaccccgg acgccttcta tacgtggccg gtgaagctgg gtgaactgac cccgcgtggc 360
ggtgaactga tcgcctatct gggtcactac tggcgtcagc gcctggtggc agatggtctg 420
ctgccgaaaa agggctgccc gcagagcggt caagttgcaa ttatcgctga tgtcgacgaa 480
cgtacccgca aaacgggtga agcatttgcg gccggtctgg caccggattg cgccattacc 540
gttcatacgc aggcagatac cagctctccg gacccgctgt tcaacccgct gaaaaccggc 600
gtctgtcagc tggatgtcgc gcaagtgacg gacgccattc tggaacgtgc aggcggttcc 660
atcgctgatt ttaccggtca ctaccagacg gcattccgtg aactggaacg cgttctgaac 720
tttccgcagt caaatctggc gctgaaacgc gaaaagcagg atgaaagtgc gtccctgacc 780
caagccctgc cgagtgaact gaaagtctcc gccgacaatg tgtcactgac cggcgcatgg 840
tcactggctt cgatgctgac ggaaattttt ctgctgcagc aagcacaggg tatgccggaa 900
ccgcagttcg acctgctgca acgtaccccg gaagtggcac gttcgcgcgc cacgccgctg 960
ctggatctga ttaaaaccgc tctgacgccg catccgccgc agaagcaagc gtatggcgtg 1020
accctgccga cgagcgttct gtttatcgcg ggtcacgaca ccaacctggc aaatctgggc 1080
ggtgctctgg aactgcagtg gaccctgccg ggtcaaccgg ataacacgcc gccgggcggt 1140
gaactggttt tcgaacgttg gcgtcgcctg agcgacaatt ctcagtggat ccaagttagc 1200
ctggtctttc agaccctgca gcaaatgcgc gataaaaccc cgctgttcct gaacacgccg 1260
ccgggcgaag tgaagctgac cctggcgggt tgcgaagaac gtaacgccca gggcatgtgt 1320
tctctggcag gttttaccca gattgttaat gaagcacgca tcccggcttg tagtctgggg 1380
ggcgcagaag cagctgccaa agaggcggcc gcaaaggtca atctgtgtct ggacctgaaa 1440
acgcaagtgc aaaccccgca aggcatgaag gaaatctcaa acatccaagt cggtgacctg 1500
gtgctgtcga ataccggcta taacgaagtg ctgaatgttt ttccgaagag caaaaagaaa 1560
tcttacaaga tcacgctgga agatggcaag gaaattattt gcagcgaaga acatctgttc 1620
ccgacccaga cgggcgaaat gaatatctcc ggcggtctga aagaaggcat gtgtctgtac 1680
gtcaaggaat aa 1692
<210> 213
<211> 583
<212> PRT
<213> Artificial Sequence
<400> 213
Met Thr Lys Lys Ile Thr Lys Ile Glu Glu Leu Asp Glu Arg Glu Leu
1 5 10 15
Ile Asp Ile Glu Val Ser Gly Asn His Leu Phe Tyr Ala Asn Asp Ile
20 25 30
Gly Thr His Ala Ser Ala Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala
35 40 45
Lys Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Ala Leu Asn Thr Pro
50 55 60
Gln Ser Ala Phe Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val
65 70 75 80
Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe Thr Gln
85 90 95
Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro Val Lys
100 105 110
Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu Gly
115 120 125
His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys Lys
130 135 140
Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp Glu
145 150 155 160
Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro Asp
165 170 175
Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro Asp Pro
180 185 190
Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val Ala Gln
195 200 205
Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp Phe
210 215 220
Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu Asn
225 230 235 240
Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp Glu Ser
245 250 255
Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala Asp
260 265 270
Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu Thr Glu
275 280 285
Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp Gly
290 295 300
Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu His Asn
305 310 315 320
Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser Arg
325 330 335
Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His Pro
340 345 350
Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu Phe
355 360 365
Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu Glu
370 375 380
Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly
385 390 395 400
Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln Trp
405 410 415
Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp Lys
420 425 430
Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr Leu
435 440 445
Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala Gly
450 455 460
Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu Gly
465 470 475 480
Gly Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Val Asn Leu Cys
485 490 495
Leu Asp Leu Lys Thr Gln Val Gln Thr Pro Gln Gly Met Lys Glu Ile
500 505 510
Ser Asn Ile Gln Val Gly Asp Leu Val Leu Ser Asn Thr Gly Tyr Asn
515 520 525
Glu Val Leu Asn Val Phe Pro Lys Ser Lys Lys Lys Ser Tyr Lys Ile
530 535 540
Thr Leu Glu Asp Gly Lys Glu Ile Ile Cys Ser Glu Glu His Leu Phe
545 550 555 560
Pro Thr Gln Thr Gly Glu Met Asn Ile Ser Gly Gly Leu Lys Glu Gly
565 570 575
Met Cys Leu Tyr Val Lys Glu
580
<210> 214
<211> 1692
<212> DNA
<213> Artificial Sequence
<400> 214
atgacgaaga aaattacgaa gatcgaagaa ctggatgaac gtgaactgat tgacatcgaa 60
gttagcggca accatctgtt ttacgcgaat gacattggga cccacaacgc cgcagccgaa 120
gccgctgcga aggaggcagc tgcgaaagaa gcggctgcaa aagaagcggc agctaaggct 180
ttgaataccc cgcaatcggc tttcgctcaa tcggaaccgg aactgaaact ggaaagtgtg 240
gttattgtgt ctcgtcatgg cgttcgcgct ccgaccaaat ttacgcagct gatgcaagat 300
gtcaccccgg acgccttcta tacgtggccg gtgaagctgg gtgaactgac cccgcgtggc 360
ggtgaactga tcgcctatct gggtcactac tggcgtcagc gcctggtggc agatggtctg 420
ctgccgaaaa agggctgccc gcagagcggt caagttgcaa ttatcgctga tgtcgacgaa 480
cgtacccgca aaacgggtga agcatttgcg gccggtctgg caccggattg cgccattacc 540
gttcatacgc aggcagatac cagctctccg gacccgctgt tcaacccgct gaaaaccggc 600
gtctgtcagc tggatgtcgc gcaagtgacg gacgccattc tggaacgtgc aggcggttcc 660
atcgctgatt ttaccggtca ctaccagacg gcattccgtg aactggaacg cgttctgaac 720
tttccgcagt caaatctggc gctgaaacgc gaaaagcagg atgaaagtgc gtccctgacc 780
caagccctgc cgagtgaact gaaagtctcc gccgacaatg tgtcactgac cggcgcatgg 840
tcactggctt cgatgctgac ggaaattttt ctgctgcagc aagcacaggg tatgccggaa 900
ccgcagttcg acctgctgca acgtaccccg gaagtggcac gttcgcgcgc cacgccgctg 960
ctggatctga ttaaaaccgc tctgacgccg catccgccgc agaagcaagc gtatggcgtg 1020
accctgccga cgagcgttct gtttatcgcg ggtcacgaca ccaacctggc aaatctgggc 1080
ggtgctctgg aactgcagtg gaccctgccg ggtcaaccgg ataacacgcc gccgggcggt 1140
gaactggttt tcgaacgttg gcgtcgcctg agcgacaatt ctcagtggat ccaagttagc 1200
ctggtctttc agaccctgca gcaaatgcgc gataaaaccc cgctgttcct gaacacgccg 1260
ccgggcgaag tgaagctgac cctggcgggt tgcgaagaac gtaacgccca gggcatgtgt 1320
tctctggcag gttttaccca gattgttaat gaagcacgca tcccggcttg tagtctgggg 1380
ggcgcagaag cagctgccaa agaggcggcc gcaaaggtca atctgtgtct ggacctgaaa 1440
acgcaagtgc aaaccccgca aggcatgaag gaaatctcaa acatccaagt cggtgacctg 1500
gtgctgtcga ataccggcta taacgaagtg ctgaatgttt ttccgaagag caaaaagaaa 1560
tcttacaaga tcacgctgga agatggcaag gaaattattt gcagcgaaga acatctgttc 1620
ccgacccaga cgggcgaaat gaatatctcc ggcggtctga aagaaggcat gtgtctgtac 1680
gtcaaggaat aa 1692
<210> 215
<211> 583
<212> PRT
<213> Artificial Sequence
<400> 215
Met Thr Lys Lys Ile Thr Lys Ile Glu Glu Leu Asp Glu Arg Glu Leu
1 5 10 15
Ile Asp Ile Glu Val Ser Gly Asn His Leu Phe Tyr Ala Asn Asp Ile
20 25 30
Gly Thr His Asn Ala Ala Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala
35 40 45
Lys Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Ala Leu Asn Thr Pro
50 55 60
Gln Ser Ala Phe Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val
65 70 75 80
Val Ile Val Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe Thr Gln
85 90 95
Leu Met Gln Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro Val Lys
100 105 110
Leu Gly Glu Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu Gly
115 120 125
His Tyr Trp Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys Lys
130 135 140
Gly Cys Pro Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp Glu
145 150 155 160
Arg Thr Arg Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro Asp
165 170 175
Cys Ala Ile Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro Asp Pro
180 185 190
Leu Phe Asn Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val Ala Gln
195 200 205
Val Thr Asp Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp Phe
210 215 220
Thr Gly His Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu Asn
225 230 235 240
Phe Pro Gln Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp Glu Ser
245 250 255
Ala Ser Leu Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala Asp
260 265 270
Asn Val Ser Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu Thr Glu
275 280 285
Ile Phe Leu Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp Gly
290 295 300
Arg Ile Thr Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu His Asn
305 310 315 320
Ala Gln Phe Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser Arg
325 330 335
Ala Thr Pro Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His Pro
340 345 350
Pro Gln Lys Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu Phe
355 360 365
Ile Ala Gly His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu Glu
370 375 380
Leu Gln Trp Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly
385 390 395 400
Glu Leu Val Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln Trp
405 410 415
Ile Gln Val Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp Lys
420 425 430
Thr Pro Leu Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr Leu
435 440 445
Ala Gly Cys Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala Gly
450 455 460
Phe Thr Gln Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu Gly
465 470 475 480
Gly Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Val Asn Leu Cys
485 490 495
Leu Asp Leu Lys Thr Gln Val Gln Thr Pro Gln Gly Met Lys Glu Ile
500 505 510
Ser Asn Ile Gln Val Gly Asp Leu Val Leu Ser Asn Thr Gly Tyr Asn
515 520 525
Glu Val Leu Asn Val Phe Pro Lys Ser Lys Lys Lys Ser Tyr Lys Ile
530 535 540
Thr Leu Glu Asp Gly Lys Glu Ile Ile Cys Ser Glu Glu His Leu Phe
545 550 555 560
Pro Thr Gln Thr Gly Glu Met Asn Ile Ser Gly Gly Leu Lys Glu Gly
565 570 575
Met Cys Leu Tyr Val Lys Glu
580
<210> 216
<211> 13
<212> PRT
<213> Artificial Sequence
<400> 216
Ala His Ile Val Met Val Asp Ala Tyr Lys Pro Thr Lys
1 5 10
<210> 217
<211> 13
<212> PRT
<213> Artificial Sequence
<400> 217
Ala His Ile Val Met Val Ala Ala Tyr Lys Pro Thr Lys
1 5 10
<210> 218
<211> 1236
<212> DNA
<213> Artificial Sequence
<400> 218
gctgcgcagt ccgagccgga gctgaagctg gagtccgtgg tgatcgtgtc gcgccacggg 60
gtgcgcgccc cgaccaagtt cacgcagctc atgcaggacg tgaccccgga cgccttctac 120
acctggccgg tgaagctcgg cgagctgacc ccgcgcggcg gcgagctgat cgcctacctc 180
ggccactact ggcgccagcg cctcgtggcc gacggcctcc tcccgaagaa gggctgcccg 240
cagtccggcc aggtggcgat catcgccgac gtggacgagc gcacccgcaa gacgggcgag 300
gccttcgccg ccggcctcgc cccggactgc gccatcaccg tgcacaccca ggccgacacc 360
tcctccccgg acccgctctt caacccgctc aagaccggcg tgtgccagct cgacgtggcc 420
caggtgaccg acgccatcct ggagcgcgcc ggcggctcca tcgccgactt caccggccac 480
taccagaccg ccttccgcga gctggagcgc gtgctcaact tcccgcagtc gaacctcgcc 540
ctcaagcgcg agaagcagga cgagtccgcc tccctcaccc aggccctccc gtccgagctg 600
aaggtgtccg ccgacaacgt gtccctcacc ggcgcctggt ccctcgcctc catgctcacc 660
gaaatcttcc tcctccagca ggcccagggc atgccggagc cgggctgggg ccgcatcacc 720
gactcccacc agtggaacac cctcctctcc ctccacaacg cccagttcga cctcctccag 780
cgcaccccgg aggtggcccg ctcccgcgcc accccgctcc tcgacctcat caagaccgcc 840
ctcaccccgc acccgccgca gaagcaggcc tacggcgtga ccctcccgac ctcggtgctc 900
ttcatcgccg gccacgacac caacctcgcc aacctcggcg gcgccctgga gctgcagtgg 960
accctcccgg gccagccgga caacaccccg ccgggcggcg agctggtgtt cgagcgctgg 1020
cgccgcctct ccgacaactc ccagtggatt caggtgtccc tcgtgttcca gaccctccag 1080
cagatgcgcg acaagacccc gctcttcctc aacaccccgc cgggcgaggt gaagctcacc 1140
ctggccggct gcgaggagcg caacgcgcag ggcatgtgct ccctcgccgg cttcacccag 1200
atcgtgaacg aggcccgcat cccggcctgc tccctc 1236
<210> 219
<211> 412
<212> PRT
<213> Artificial Sequence
<400> 219
Ala Ala Gln Ser Glu Pro Glu Leu Lys Leu Glu Ser Val Val Ile Val
1 5 10 15
Ser Arg His Gly Val Arg Ala Pro Thr Lys Phe Thr Gln Leu Met Gln
20 25 30
Asp Val Thr Pro Asp Ala Phe Tyr Thr Trp Pro Val Lys Leu Gly Glu
35 40 45
Leu Thr Pro Arg Gly Gly Glu Leu Ile Ala Tyr Leu Gly His Tyr Trp
50 55 60
Arg Gln Arg Leu Val Ala Asp Gly Leu Leu Pro Lys Lys Gly Cys Pro
65 70 75 80
Gln Ser Gly Gln Val Ala Ile Ile Ala Asp Val Asp Glu Arg Thr Arg
85 90 95
Lys Thr Gly Glu Ala Phe Ala Ala Gly Leu Ala Pro Asp Cys Ala Ile
100 105 110
Thr Val His Thr Gln Ala Asp Thr Ser Ser Pro Asp Pro Leu Phe Asn
115 120 125
Pro Leu Lys Thr Gly Val Cys Gln Leu Asp Val Ala Gln Val Thr Asp
130 135 140
Ala Ile Leu Glu Arg Ala Gly Gly Ser Ile Ala Asp Phe Thr Gly His
145 150 155 160
Tyr Gln Thr Ala Phe Arg Glu Leu Glu Arg Val Leu Asn Phe Pro Gln
165 170 175
Ser Asn Leu Ala Leu Lys Arg Glu Lys Gln Asp Glu Ser Ala Ser Leu
180 185 190
Thr Gln Ala Leu Pro Ser Glu Leu Lys Val Ser Ala Asp Asn Val Ser
195 200 205
Leu Thr Gly Ala Trp Ser Leu Ala Ser Met Leu Thr Glu Ile Phe Leu
210 215 220
Leu Gln Gln Ala Gln Gly Met Pro Glu Pro Gly Trp Gly Arg Ile Thr
225 230 235 240
Asp Ser His Gln Trp Asn Thr Leu Leu Ser Leu His Asn Ala Gln Phe
245 250 255
Asp Leu Leu Gln Arg Thr Pro Glu Val Ala Arg Ser Arg Ala Thr Pro
260 265 270
Leu Leu Asp Leu Ile Lys Thr Ala Leu Thr Pro His Pro Pro Gln Lys
275 280 285
Gln Ala Tyr Gly Val Thr Leu Pro Thr Ser Val Leu Phe Ile Ala Gly
290 295 300
His Asp Thr Asn Leu Ala Asn Leu Gly Gly Ala Leu Glu Leu Gln Trp
305 310 315 320
Thr Leu Pro Gly Gln Pro Asp Asn Thr Pro Pro Gly Gly Glu Leu Val
325 330 335
Phe Glu Arg Trp Arg Arg Leu Ser Asp Asn Ser Gln Trp Ile Gln Val
340 345 350
Ser Leu Val Phe Gln Thr Leu Gln Gln Met Arg Asp Lys Thr Pro Leu
355 360 365
Phe Leu Asn Thr Pro Pro Gly Glu Val Lys Leu Thr Leu Ala Gly Cys
370 375 380
Glu Glu Arg Asn Ala Gln Gly Met Cys Ser Leu Ala Gly Phe Thr Gln
385 390 395 400
Ile Val Asn Glu Ala Arg Ile Pro Ala Cys Ser Leu
405 410

Claims (58)

1.一种工程化植酸酶,包括目标植酸酶、第一结合元件和第二结合元件,其中,所述第一结合元件和所述第二结合元件中的每个均与所述目标植酸酶融合,所述第一结合元件与所述第二结合元件相互作用以引起所述工程化植酸酶的环化,并且增强所述目标植酸酶的热稳定性,并且其中,所述第一结合元件或所述第二结合元件选自由以下各项组成的组中:标签结构域、捕捉结构域、内含肽或其部分和卷曲螺旋二聚化结构域或其部分。
2.根据权利要求1所述的工程化植酸酶,其中,在所述相互作用时,所述第一结合元件和所述第二结合元件中的每个都能够自发地从所述工程化植酸酶释放出来。
3.根据权利要求1所述的工程化植酸酶,其中,所述第一结合元件或所述第二结合元件与所述目标植酸酶的N-末端或C-末端融合。
4.根据权利要求3所述的工程化植酸酶,其中,所述第一结合元件的C末端连接至所述目标植酸酶的N末端,并与其邻接。
5.根据权利要求3所述的工程化植酸酶,其中,所述第二结合元件的N末端连接至所述目标植酸酶的C末端,并与其邻接。
6.根据权利要求1所述的工程化植酸酶,其中,所述目标植酸酶选自由以下各项组成的组中:来源于大肠杆菌(Escherichia coli)、黑曲霉(Aspergillus niger)、隔孢伏革菌(Peniophora lycii)、粗糙脉孢菌(Neurospora crassa)和意大利拟牛链球菌(Schwaniomyces accidentalis)的植酸酶。
7.根据权利要求1所述的工程化植酸酶,其中,所述目标植酸酶包括与选自由SEQ IDNO:53-54、56和219组成的组中的参考序列具有至少90%同一性的氨基酸序列。
8.根据权利要求1所述的工程化植酸酶,其中,所述第一结合元件是内含肽的C-内含肽。
9.根据权利要求8所述的工程化植酸酶,其中,所述C-内含肽包括与选自由以下序列组成的组中的参考序列具有至少90%同一性的氨基酸序列:SEQ ID NO:2、4、6、8、10、12、14、16、18、20、22、24、26、28、30、32、189、191和195。
10.根据权利要求1所述的工程化植酸酶,其中,所述第二结合元件是内含肽的N-内含肽。
11.根据权利要求10所述的工程化植酸酶,其中,所述N-内含肽包括与选自由以下序列组成的组中的参考序列具有至少90%同一性的氨基酸序列:SEQ ID NO:1、3、5、7、9、11、13、15、17、19、21、23、25、27、29、31、187和193。
12.根据权利要求1所述的工程化植酸酶,其中,所述第一结合元件是所述卷曲螺旋二聚化结构域的C-螺旋。
13.根据权利要求12所述的工程化植酸酶,其中,所述C-螺旋包括与SEQ ID NO:38或40的参考序列具有至少90%同一性的氨基酸序列。
14.根据权利要求1所述的工程化植酸酶,其中,所述第二结合元件是卷曲螺旋二聚化结构域的N-螺旋。
15.根据权利要求14的工程化植酸酶,其中,所述N-螺旋包括与SEQ ID NO:37或39的参考序列具有至少90%同一性的氨基酸序列。
16.根据权利要求1所述的工程化植酸酶,其中,所述第一结合元件是标签结构域。
17.根据权利要求16所述的工程化植酸酶,其中,所述标签结构域包括与SEQ ID NO:33或34的参考序列具有至少90%同一性的氨基酸序列。
18.根据权利要求1所述的工程化植酸酶,其中,所述第二结合元件是捕捉结构域。
19.根据权利要求18所述的工程化植酸酶,其中,所述捕捉结构域包括与SEQ ID NO:35或36的参考序列具有至少90%同一性的氨基酸序列。
20.根据权利要求1所述的工程化植酸酶,其中,还包括第一连接子,其中,所述第一连接子与所述第一结合元件和所述目标植酸酶邻接,并在它们之间。
21.根据权利要求20所述的工程化植酸酶,其中,所述第一连接子包括与选自由以下序列组成的组中的序列具有至少90%同一性的序列:SEQ ID NO:41、43、45、47、48、50和51。
22.根据权利要求1所述的工程化植酸酶,其中,还包括第二连接子,其中,所述第二连接子与所述目标植酸酶和所述第二结合元件邻接,并在它们之间。
23.根据权利要求22所述的工程化植酸酶,其中,所述第二连接子包括与选自由以下序列组成的组中的序列具有至少90%同一性的序列:SEQ ID NO:42、44、46、49、50和51。
24.根据权利要求1所述的工程化植酸酶,其中,包括与选自由以下序列组成的组中的参考序列具有至少90%同一性的氨基酸序列:SEQ ID NO:58、60、62、64、66、68、70、72、74、76、78、80、82、84、86、88、90、92、93、95、97、99、101、103、105、107、109、111、113、115、117、119、201、203、205和207。
25.根据权利要求1所述的工程化植酸酶,其中,所述植酸酶的活性在70℃至90℃范围的温度下是稳定的。
26.根据权利要求1所述的工程化植酸酶,其中,所述工程化植酸酶包括在宿主中,其中,所述宿主选自由微生物、植物细胞、噬菌体、病毒、哺乳动物细胞和昆虫细胞组成的组中。
27.一种编码权利要求1-26中任意一项所述的工程化植酸酶的工程化核酸。
28.一种编码工程化植酸酶的工程化核酸,其中,所述工程化植酸酶包括目标植酸酶、第一结合元件和第二结合元件,其中,所述第一结合元件和所述第二结合元件中的每个均与所述目标植酸酶融合,所述第一结合元件与所述第二结合元件相互作用以引起所述工程化植酸酶的环化,并且增强所述目标植酸酶的热稳定性,并且其中,所述第一结合元件或所述第二结合元件选自由以下各项组成的组中:标签结构域、捕捉结构域、内含肽或其部分以及卷曲螺旋二聚化结构域或其部分。
29.根据权利要求28所述的工程化核酸,其中,在所述相互作用时,所述第一结合元件和所述第二结合元件中的每个都能够自发地从所述工程化植酸酶释放出来。
30.根据权利要求28所述的工程化核酸,其中,包括编码选自由以下各项组成的组中的目标植酸酶的序列:来源于大肠杆菌(Escherichia coli)、黑曲霉(Aspergillus niger)、隔孢伏革菌(Peniophora lycii)、粗糙脉孢菌(Neurospora crassa)和意大利拟牛链球菌(Schwaniomyces accidentalis)的植酸酶。
31.根据权利要求28所述的工程化核酸,其中,包括编码所述目标植酸酶的序列,并且该序列与选自由SEQ ID NO:52、55、185和219组成的组中的参考序列具有至少90%的同一性。
32.根据权利要求28所述的工程化核酸,其中,所述第一结合元件与所述目标植酸酶的N-末端融合。
33.根据权利要求28所述的工程化核酸,其中,所述第二结合元件与所述目标植酸酶的C-末端融合。
34.根据权利要求28所述的工程化核酸,其中,包括编码所述第一结合元件的序列,其中,所述第一结合元件是内含肽的C-内含肽。
35.根据权利要求34所述的工程化核酸,其中,包括与选自由以下序列组成的组中的参考序列具有至少90%同一性的序列:SEQ ID NO:143、145、147、149、151、153、155、157、159、161、163、165、167、169、171、173、188、190和194。
36.根据权利要求28所述的工程化核酸,其中,包括编码所述第二结合元件的序列,其中,所述第二结合元件是内含肽的N-内含肽。
37.根据权利要求36所述的工程化核酸,其中,包括与选自由以下序列组成的组中的参考序列具有至少90%同一性的序列:SEQ ID NO:142、144、146、148、150、152、154、156、158,160、162、164、166、168、170、172、186和192。
38.根据权利要求28所述的工程化核酸,其中,包括编码所述第一结合元件的序列,其中,所述第一结合元件是卷曲螺旋二聚化结构域的C-螺旋。
39.根据权利要求38所述的工程化核酸,其中,包括与SEQ ID NO:179或181的参考序列具有至少90%同一性的序列。
40.根据权利要求28所述的工程化核酸,其中,包括编码所述第二结合元件的序列,其中,所述第二结合元件是卷曲螺旋二聚化结构域的N-螺旋。
41.根据权利要求40所述的工程化核酸,其中,包括与SEQ ID NO:178或180的参考序列具有至少90%同一性的序列。
42.根据权利要求28所述的工程化核酸,其中,包括编码所述第一结合元件的序列,其中,所述第一结合元件是标签结构域。
43.根据权利要求42所述的工程化核酸,其中,包括与SEQ ID NO:174或175的参考序列具有至少90%同一性的序列。
44.根据权利要求28所述的工程化核酸,其中,包括编码所述第二结合元件的序列,其中,所述第二结合元件是捕捉结构域。
45.根据权利要求44所述的工程化核酸,其中,包括与SEQ ID NO:176或177的参考序列具有至少90%同一性的序列。
46.根据权利要求28所述的工程化核酸,其中,还包括编码第一连接子的序列,其中,所述第一连接子与所述第一结合元件和所述目标植酸酶邻接,并在它们之间。
47.根据权利要求46所述的工程化核酸,其中,包括与选自由SEQ ID NO:120、122、124、126、182、183和184组成的组中的序列具有至少90%同一性的序列。
48.根据权利要求28所述的工程化核酸,其中,还包括编码第二连接子的序列,其中,所述第二连接子与所述目标植酸酶和所述第二结合元件邻接,并在它们之间。
49.根据权利要求48所述的工程化核酸,其中,包括与选自由SEQ ID NO:121、123、125、127、183和184组成的组中的序列具有至少90%同一性的序列。
50.根据权利要求28所述的工程化核酸,其中,包括与选自由以下序列组成的组中的参考序列具有至少90%同一性的序列:SEQ ID NO:57、59、61、63、65、67、69、71、73、75、77、79、81、83、85、87、89、91、94、96、98、100、102、104、106、108、110、112、114、116、118、128-133、200、202、204和206。
51.根据权利要求28所述的工程化核酸,其中,包括编码所述工程化植酸酶的序列,所述工程化植酸酶在70℃至90℃范围的温度下具有稳定的植酸酶活性。
52.根据权利要求28所述的工程化核酸,其中,所述工程化核酸在宿主中表达,其中,所述宿主选自由以下各项组成的组中:微生物、植物细胞、噬菌体、病毒、哺乳动物细胞和昆虫细胞。
53.一种包括权利要求28-52中任意一项所述的工程化核酸的载体。
54.一种包括权利要求1-26中任意一项所述的工程化植酸酶的宿主,其中,所述宿主选自由以下各项组成的组中:微生物、植物细胞、噬菌体、病毒、哺乳动物细胞和昆虫细胞。
55.一种增强目标植酸酶的热稳定性的方法,包括生产权利要求1-26中任意一项所述的工程化植酸酶。
56.一种包括工程化植酸酶的动物饲料,所述工程化植酸酶包括目标植酸酶、第一结合元件和第二结合元件,其中,所述第一结合元件和所述第二结合元件中的每个与所述目标植酸酶融合,所述第一结合元件与所述第二结合元件相互作用以引起所述工程植酸酶的环化,并且增强所述目标植酸酶的热稳定性,并且其中,所述第一结合元件或所述第二结合元件选自由以下各项组成的组中:标签结构域、捕捉结构域、内含肽或其部分和卷曲螺旋二聚化结构域或其部分。
57.一种包括权利要求1-26中任意一项所述的工程化植酸酶的动物饲料。
58.一种制备动物饲料的方法,包括将权利要求1-26中任意一项所述的工程化植酸酶添加至所述动物饲料中。
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Families Citing this family (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014197547A1 (en) 2013-06-04 2014-12-11 Tribiotica Llc Methods and compositions for templated assembly of nucleic acid specific heterocompounds
EP3227476B1 (en) 2014-12-02 2021-02-24 Tribiotica Llc Methods and kits for theranostic applications
CN108026145B (zh) * 2015-09-18 2022-06-10 谷万达公司 工程化植酸酶及其使用方法
US11253536B2 (en) 2016-11-21 2022-02-22 Tribiotica Llc Methods for directed folding assembly or dimerization of proteins by templated assembly reactions
WO2018094070A1 (en) 2016-11-21 2018-05-24 Tribiotica Llc Methods for preventing titration of bimolecular templated assembly reactions by structurally-determined differential hybridizations
GB201706430D0 (en) 2017-04-24 2017-06-07 Univ Oxford Innovation Ltd Proteins and peptide tags with enhanced rate of spontaneous isopeptide bond formation and uses thereof
JP2020535114A (ja) * 2017-08-11 2020-12-03 トリビオティカ・エルエルシー テンプレート化アセンブリによって認識分子に結合するエピトープの生成方法
US20210238578A1 (en) * 2018-04-26 2021-08-05 Danisco Us Inc Inorganic phosphate as a stabilizer for phytase enzymes
CN110872585B (zh) * 2018-08-29 2021-10-19 北京科技大学 用SpyTag/SpyCatcher环化的L-β-羟基-α-氨基酸合成酶及其用途
US10995325B2 (en) * 2019-03-21 2021-05-04 Fornia Biosolutions, Inc. Additional phytase variants and methods
CN110846336B (zh) * 2019-11-26 2022-12-20 温氏食品集团股份有限公司 一种多功能融合酶xaet和多功能融合酶定点整合真核特异表达载体及其构建方法
EP4119660A1 (en) * 2021-07-16 2023-01-18 AB Enzymes Oy Phytase variants
EP4119661A1 (en) * 2021-07-16 2023-01-18 AB Enzymes Oy Phytase variants with improved stability and ip4 activity
CN113832126A (zh) * 2021-11-27 2021-12-24 中国农业科学院北京畜牧兽医研究所 一种提高植酸酶热稳定性的方法及融合植酸酶
CN114672505A (zh) * 2022-03-15 2022-06-28 江苏省中国科学院植物研究所 一种功能性表达细胞色素p450酶的方法及其应用
CN115093470B (zh) * 2022-06-30 2023-03-24 广州市乾相生物科技有限公司 内含肽Mtu RecA突变体及其在生产谷胱甘肽GSH中的应用
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP3754936B2 (ja) * 2001-07-10 2006-03-15 キヤノン株式会社 ポリヒドロキシアルカノエート含有構造体およびその製造方法
WO2003056904A2 (en) 2002-01-08 2003-07-17 Raab Michael R Transgenic plants expressing civps or intein modified proteins and related method
US7872096B2 (en) 2004-05-24 2011-01-18 Rigel Pharmaceuticals, Inc. Methods for cyclizing synthetic polymers
US7629139B2 (en) 2005-06-21 2009-12-08 Ab Enzymes Gmbh Extraction methods and assays for feed enzymes
WO2008143679A2 (en) * 2006-06-01 2008-11-27 Verenium Corporation Nucleic acids and proteins and methods for making and using them
ES2438268T3 (es) 2006-09-21 2014-01-16 Verenium Corporation Fitasas, ácidos nucleicos que las codifican y métodos para su producción y uso
WO2009073399A2 (en) 2007-12-03 2009-06-11 Syngenta Participations Ag Engineering enzymatically susceptible proteins
CN101638642B (zh) 2008-08-01 2012-02-22 中国农业科学院生物技术研究所 提高了热稳定性及蛋白酶抗性的环化植酸酶
CN102575237A (zh) * 2009-10-22 2012-07-11 巴斯夫欧洲公司 合成的植酸酶变体
US10407742B2 (en) 2009-11-06 2019-09-10 Agrivida, Inc. Intein-modified enzymes, their production and industrial applications
US8420387B2 (en) 2009-11-06 2013-04-16 Agrivida, Inc. Intein-modified enzymes, their production and industrial applications
US9464333B2 (en) 2009-11-06 2016-10-11 Agrivida, Inc. Intein-modified enzymes, their production and industrial applications
US20120190609A1 (en) 2010-08-30 2012-07-26 Martin Bader Method for producing a lipid particle, the lipid particle itself and its use
ES2799500T3 (es) 2012-02-07 2020-12-18 Danisco Us Inc Método para la mejora de la estabilidad de fitasa con ácido fítico, y composiciones que comprenden fitasa y ácido fítico
WO2013127700A1 (en) 2012-02-29 2013-09-06 F. Hoffmann-La Roche Ag Method for reduction of 1->2 reading frame shifts
CN108026145B (zh) * 2015-09-18 2022-06-10 谷万达公司 工程化植酸酶及其使用方法

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