CN1882704A - 尼帕病毒检测方法和提供抗汉尼巴病毒的免疫保护的方法 - Google Patents
尼帕病毒检测方法和提供抗汉尼巴病毒的免疫保护的方法 Download PDFInfo
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Abstract
本发明提供了用于监测尼帕病毒感染的动物模型、用于定量检测及快速表征样品中的尼帕病毒RNA的方法、能用于给个体提供免疫保护作用的组合物、以及中和尼帕病毒及亨德拉病毒并能用于预防、治疗、和/或防护的单克隆抗体。
Description
本发明涉及一种针对样品中的尼帕病毒(Nipah virus)的检测方法以及一种用于提供抗尼帕病毒和亨德拉病毒(Hendra virus)感染的免疫保护的方法。
1998年尼帕病毒(Nipah virus,NiV)出现于马来西亚,造成了猪和人的严重的发病率和死亡率(Chua,2000,Science.288:1432-5)。最可能的动物源性感染包括作为天然宿主的狐蝠(Pteroid bats,Flying fox),它们通过它们的尿或残留于部分吃过的水果上的残留物而将尼帕病毒传染给猪群(Chua,et al 2002,Microbes Infect.4:145-51;Chua,K.B.2003,J.Clin.Microbiol.26:265-275)。与感染动物直接接触的猪饲养者和屠宰场工人是最易感的人群。经紧密接触而造成的猪到人的传播是最常见的污染途径,其中猪扮演着病毒的部分扩增宿主(Parashar,et al 2000,J Infect Dis.181:1755-9;Mohd Nor et al 2000,Rev Sci Tech Off Int Epiz.19(1):160-5)。被感染的猪主要患呼吸道疾病,死亡率小于5%,而在265例发生严重急性热病性脑炎综合征的人患者中,有105例死亡,并且四分之一的幸存者都有残留的神经副作用(Goh,et al 2000,New Engl J Med.342:1229-35;Chong,et al 2002,Can J Neurol Sci.29:83-7;Lee,et al 1999,AnnNeurol.46:428-32)。
尼帕病毒是一种副粘病毒科的副粘病毒亚科的成员。它的生物学性能和基因组结构将病毒以及与其紧密相关的亨德拉病毒分型到一个新的属,称为汉尼巴病毒(Henipavirus)(Wang,et al 2000,J Virology.74:9972-9979)。尼帕病毒含有约18,000个核苷酸的单链RNA,其与复制复合物(核蛋白(N)、磷蛋白(P)、和聚合酶(L))的病毒蛋白结合,单链RNA被含有附着蛋白(G)和融合蛋白(F)的脂质双层的被膜所包绕(Chua,2000,Science.288:1432-5;Wang,et al 2001,Microbes and Infection 3,279-287;Chan,et al 2001,J Gen Virol.82:2151-5)。
狐蝠(Pteropus sp.)东半球果蝠的广泛分布是从印度洋的西部岛屿向东南部延伸,跨越东南亚和澳大利亚东北部,一直到太平洋的西南岛屿。对造成汉尼巴病毒出现的潜在的因素了解得很少(Morse,S.S.1995.Emerg Infect Dis.1(1):7-15;Field,et al 2001,Microbes Infect.3:307-314)。2002年已经证实在柬埔寨出现了尼帕病毒,因为已经在果蝠中发现了抗NiV抗体(Olson,et al 2002,Emerg Infect Dis.8:987-988),并推测孟加拉国是在2001、2003年以及更最近的是在2004年(ProMed 2002 Nipah-like virus-Bangladesh(2001,2004):Archive numbers 20020830.5187-20040423.1127)(ICDDR,B 2003,Health and Science Bulletin,ISSN 1729-343X,vol.1:1-6)。如果想发展出有效的预防或治疗人体体内的尼帕病毒感染的计划,必需要确定对于诱导保护性应答是重要的病毒抗原,以及必须要制定可能的免疫预防性治疗。
发展用于正确监视汉尼巴病毒循环的特异的血清学和病毒学的诊断方法也是优先的(Daniels,et al 2001,Microbes Infect.3:289-95)。对在动物传染周期(zoonotic cycle)中的或在急性脑炎患者中的病毒的快速诊断将有助于选用医学、兽医学和环境水平上的合适的测定方法。已经开发出用于测试传染性疾病和细胞培养物中的病毒载量的依据于TaqManTM技术的实时聚合酶链式反应方法(Heid,et al 1996,Genome Res.10:986-94;Klein,et al 2003,J Virol Methods.107(2):169-75)。
就本质而言,副粘病毒可以感染人和动物。病毒常常优先地感染一种物种而在第二种物种中缓慢生长。因此,在第二种物种中缓慢生长的病毒可以被用于产生“Jenner”型疫苗。为了诱导保护性应答,以相同的方式,通过使用现代生物技术,可以从等价的动物病毒中表达出作为人致病原的病毒的抗原(Schmidt,et al 2002.J Virol.76:1089-1099;Yunus,etal 1999.Arch Virol.144:1977-1990)。在某些情况中,当副粘病毒跨越物种障碍而感染人时,它们变得毒性更强。亨德拉病毒和尼帕病毒的天然宿主可能是果蝠(Chua,K.B.,et al 2002.Microbes Infect.4:145-51;Field,H.,et al 2001.Microbes Infect.3:307-314;Yob,et al 2001.Emerg Infect Dis.7:439-441),但是1994年和1998年在澳大利亚,马感染了亨德拉病毒以及1998年在马来西亚,猪感染了尼帕病毒。在两种情况中,病毒都是在第二种动物物种中扩增并导致了人的感染。尼帕病毒在猪(超过100万头猪被屠杀)以及在人(40%死亡率)所引起的疾病的严重性已经带来了极大的经济的和社会的后果。在一些患者中试用了利巴韦林,但是只有很不显著的结果(Chong,H.T.,et al 2001.Ann Neurol.49:810-813;Snell,N.J.2001.Expert Opin Pharmacother.2:1317-13124)。没有尼帕病毒特异的抗病毒药物对抗病毒的流行,如果当未来病毒流行发生时要采取有效的措施,那么尼帕病毒特异性抗病毒药物的生产仍是优先的。
综上所述,需要提供一些手段监测与汉尼巴病毒感染相关的病理生理(例如动物模型和用于定量病毒负荷的定量方法)。也需要提供一种简单的、可靠的、特异的和敏感的用于定量检测样品中的尼帕样病毒或亨德拉样病毒的检测法。此外,因为尼帕病毒和亨德拉病毒感染所造成的固有的危险,仍需要给那些需要这些保护的对象提供治疗性或保护性免疫。因此,需要鉴定一种能复制人类疾病并用于抗病毒及疫苗试验的动物模型。此外,必需要有新的方法预防或治疗汉尼巴病毒感染。
因此,本发明提供了一种仓鼠模型,它复制了人急性尼帕病毒感染的病理和发病机理。
本发明的另一个目的也提供了一种用于定量检测并快速鉴定样品中的尼帕病毒RNA的方法。
本发明的另一个目的是一种免疫原性组合物,它包括尼帕病毒糖蛋白和一种可药用载体,此外,其中免疫原性组合物是一种疫苗。
本发明的另一个目的是一种保护个体对抗尼帕病毒感染的方法,其包括给所述个体施用足以诱导所述个体的免疫应答的量的尼帕病毒糖蛋白或编码所述糖蛋白的多核苷酸。
本发明的另一个目的是一种用于保护或治愈个体对抗尼帕病毒感染的免疫原性组合物,其包括针对尼帕病毒的附着和/或融合糖蛋白的或与汉尼巴病毒属可交叉反应的抗体。
通过结合附图说明并参照下面的详细说明有助于更好地了解本发明,由此将对本发明及其本发明的许多伴随的优点有更加完整的了解。
图1:经两种途径感染尼帕病毒的7-14周龄仓鼠的生存曲线。经腹腔内和鼻内途径杀死50%仓鼠的病毒的致死剂量(LD50)分别是每只动物270pfu和47,000pfu。
图2:急性尼帕病毒感染时的血管和实质的病理。A:肝脏大动脉表现为局灶的、透壁的纤维素样坏死以及周围的炎症。B:心肌坏死以及邻近的炎症。C:肺动脉中的多个内皮细胞的多核合胞体。D:在同一肺内的内皮细胞合胞体及血管平滑肌中都证实有病毒RNA。E:脑血管的坏死和核破碎。F:定位于脑膜血管的内皮及平滑肌中的病毒抗体。
图3:急性尼帕病毒感染时的脑部病理。A:小血管的血管炎的特点是感染神经元邻近的轻微炎症。B:局灶的实质缺血、梗死及水肿的区域。C:具有嗜酸细胞包涵体的神经元。D:病毒抗原在神经元的核、胞浆以及邻近血管炎血管的突出中的免疫定位。E:定位于室管膜内层及神经元的病毒抗原。F:证实病毒RNA存在于其胞浆内的神经元。
图4:A和B:与血管炎及血栓性血管相关的肺实质的炎症。C:肾小球肾炎的特点是血栓性栓塞、炎症以及肾小球周围的合胞体形成。D:在肾小球的小管处检测到病毒抗原。E:在覆盖肾乳头的内皮细胞中发现有病毒抗原。F:在脾白质的淋巴细胞中证实有病毒抗原。
图5:TaqManTM实时RT-PCR对尼帕病毒RNA的检测。用尼帕病毒原液中提取的尼帕病毒RNA的10倍稀释液绘制出扩增曲线。重复进行从未稀释到1/106的测试。
图6:用尼帕病毒RNA的10倍系列稀释液得到的标准曲线。曲线标绘了从图5中获得的结果中计算出的Ct值对感染病毒的最初起始量(pfu/ml)的指数值。阈值是0.289601。
图7:尼帕病毒RNA转录产物的标准曲线,显示了标绘的阈值周期Ct对尼帕病毒RNA转录产物的最初量的指数值。用不同的RNA转录产物绘制出三条扩增曲线。
图8:尼帕病毒感染和Vero细胞的合胞体形成。在感染后第1天(a)和第2天(b)处理经0.01MOI感染的细胞,并用免疫荧光检测病毒抗原的存在。通过含有大量核的细胞合胞体的形成来观察细胞致病作用。用碘化丙啶(propidium iodide)染色核。
图9:在感染后第1、2、3和4天,用噬斑检测法和实时RT-PCR检测法评价在感染细胞上清液中检测到的尼帕病毒以及尼帕病毒RNA的数目。
图10:对受表达NiV G或F糖蛋白的疫苗病毒(VV)重组体感染的HeLa细胞的FACScan分析。用MOI为0.1pfu/细胞的VV-NiV.G或F或对照VV感染HeLa细胞16个小时,并用抗G或F糖蛋白的多克隆的单特异性抗血清测定细胞表面上的糖蛋白的表达。
图11:用共表达尼帕病毒G和F糖蛋白诱导融合。如图10用表达G或F糖蛋白的VV-NiV重组体感染或用两者双重感染HeLa细胞。然后用免疫荧光法检查细胞的病毒表达并且也检查融合诱导。
图12:通过用表达尼帕病毒G和/或F糖蛋白的VV重组体免疫接种保护仓鼠对抗尼帕病毒的致死性攻击。用VV.NiV G或F或两者间隔1个月免疫接种仓鼠两次,并在末次免疫接种3个月后用尼帕病毒攻击(7-8只动物/组)。每天检查动物。
图13:在用VV重组体免疫接种和用尼帕病毒攻击之后的抗体应答。在免疫接种后以及也在尼帕病毒攻击后一段时间内,给仓鼠抽血。用(A)中和法和用(B)ELISA检测抗体水平。
图14:仓鼠抗致死性尼帕病毒感染的被动保护。在仓鼠中生成了抗表达G或F的VV重组体的抗体,并在用尼帕病毒攻击之前2个小时,i.p.接种(0.2ml/动物)所富集的抗个别分糖蛋白或抗相等的每种糖蛋白的混和物的血清。24个小时之后给予第二次抗血清的接种(0.2ml)。用尼帕病毒攻击动物并观察43天。
图15:在被动施用多克隆的单特异性抗尼帕病毒血清时,用尼帕病毒攻击的仓鼠的免疫应答。间断地给图14中的仓鼠抽血,并用ELISA检查血清中的抗尼帕病毒抗体。
除非有特异的定义,在此所用的所有技术和科学术语具有与如分子生物学领域的普通技术人员通常所理解的相同的含义。
本发明第一次提供了实例,即金仓鼠可以被经鼻内或腹腔内途径注射的尼帕病毒感染并且死于尼帕病毒在被感染人中的特征性的脑炎综合征。此外,在尸解中所观察到的病变表现出与那些在人体组织样品中观察到的相似的病理。具体地,病变表现出病毒嗜血管内皮细胞性(形成合胞体),并导致血管炎、血栓形成、缺血、梗死、和血管周围炎症,与在人体感染中观察到的情况相似(Wong et al.,Am.J.Pathol.2002.161:2153-2167)。也已经证实中枢神经细胞的神经元是尼帕病毒的靶细胞。病毒抗原和RNA定位于血管和血管外组织,包括神经元、肺、肾脏、和脾脏。最后,从感染动物的尿液中可分离到了病毒,提供了一种非侵入性操作随诊存在病毒复制的相关方法。
因此,在本发明的一个实施方式中,提供了一种汉尼巴病毒感染的金仓鼠模型,其中用至少一种汉尼巴病毒例如尼帕病毒和亨德拉病毒感染仓鼠。这个金仓鼠模型复制出了在被感染人体中所观察到的大部分症状(即超过50%)。本模型可便利地用作一种人和非人的灵长类动物的替代物,用于诊断、病毒生产、病毒表型识别、和治疗学及预防学的评估。
本发明也第一次提供了一种多能的、可靠的、和敏感的检测法,以便快速地定量细胞培养物和生物学样品中的尼帕病毒RNA。在RNA提取过程期间,对病毒传染性的灭活容许任何参与监视和诊断该病毒的实验室检测尼帕病毒在传染流行区中的流行情况。该技术也可以涉及定量组织标本中的病毒RNA分子。已经描述了尼帕病毒可以长时间停留在人体内并引起迟发性脑炎,或它可以在最初发病数个月后复发引起复染性脑炎(Tan,et al 2002,Ann Neurol.51:703-8)。尽管在这些晚期阶段不能从脑脊液中分离出活病毒,但是通过证实脑中的病毒抗原可以揭示尼帕病毒的存在。
包括尼帕病毒和亨德拉病毒的副粘病毒在病毒表面上都具有两种糖蛋白G和F。G糖蛋白造成与细胞受体的附着,而F糖蛋白诱导了病毒和细胞膜之间的融合。G和F协同作用引起融合。本发明者已经证实了表达尼帕病毒蛋白的疫苗只有在共感染(即G+F)时才诱导融合。如果抗体要阻断感染,那么推测它们应当阻断G与其受体的附着或抑制F融合病毒被膜与细胞膜的功能。来自用各种VV重组体免疫接种的仓鼠的血清诱导了高抗体水平,却是相当低的中和抗体。在其他副粘病毒中,对附着蛋白的诱导常常是更明显的,但是本发明者发现对尼帕病毒F和尼帕病毒G的抗体应答是相同级别的,在小鼠中进行了确认研究(Tamin,et al 2002.Virology.296:190-200)。
本发明所含的基础科学技术是已知的。见,例如Sambrook et al.,Molecular Cloning:A Laboratory Manual,Third Edition,Cold Spring HarborLaboratory,New York(1999),并在此引用各种参考文献。
“分离的”指的是一种与其天然环境分离开的材料,即多核苷酸。
“重组体”指的是一种通过切割多核苷酸并将特异多核苷酸片段剪切在一起而在体外制备的、遗传加工的多核苷酸或多肽。
“多核苷酸”通常涉及多核糖核苷酸和多脱氧核糖核苷酸,也可能涉及那些非修饰的RNA或DNA或修饰的RNA或DNA。
“多肽”被理解为意思是肽或蛋白,它包括两个或多个经肽键结合的氨基酸。
“抑制”在此包括汉尼巴病毒例如尼帕病毒和/或亨德拉病毒在对象患者中的传染性的任何可测定的可重复的减低。
术语“表达载体”指的是一种编码本发明的肽并为其在所选的宿主细胞中的表达提供必需序列的多核苷酸。表达载体通常将包括一个转录启动子和终止子,或将提供在邻近于内源性启动子的整合。表达载体可以是质粒,还包括复制起点和一个或多个选择标记物。另外,表达载体可以是设计用于感染宿主的病毒重组体,或设计用于整合到宿主基因组内的优选位点上的整合载体。病毒重组体的实例是腺伴随病毒(AAV)、腺病毒、疱疹病毒、痘病毒、逆转录病毒、痘苗病毒和其他本领域已知的RNA或DNA病毒表达载体。在一个优选的实施方式中,表达载体是一种病毒载体,以及在一个特别优选的实施方式中,病毒载体是一种重组痘苗病毒。
本发明的检测方法可以采用逆转录酶-聚合酶链反应(RT-PCR),其中联合逆转录实施PCR。通常地,利用标准技术从样品组织中提取RNA,并将其逆转录生成cDNA分子。然后该cDNA被用作随后的聚合酶链反应的模板。
一旦引物和模板已经退火,用DNA聚合酶延伸引物,然后合成出模板的拷贝。然后将DNA链变性并重复该过程多次,直到生成了足够多的DNA以便容许观察,观察使用了附着于至少一种引物的荧光、放射性核素、或其他可检测部分,或其他观察所扩增的多核苷酸分子的方法例如溴化乙锭染色或分光光度法。
在这些检测法中所用的生物样品包括血液、血清、尿液、组织活检物、淋巴结、腹水、脑脊液和前列腺分泌物、以及其他组织、匀浆、和它们的提取物。利用任何标准的技术可以制备这些生物样品。
在上面所提供的方法中可以使用编码尼帕病毒和亨德拉病毒蛋白(或它们的一部分或其他变体)的多核苷酸或与该多核苷酸互补的多核苷酸。多核苷酸可以是单链的(编码的或反义的)或双链的,以及可以是DNA(cDNA或合成的)或RNA分子。附加的编码或非编码的序列可以存在于本发明的多核苷酸内,但这不是必需的,以及多核苷酸可以与其他分子和/或支持材料连接,但这并不是必需的。
利用多种技术中的任何一种技术可以制备多核苷酸。例如,可以用聚合酶链反应(PCR)从cDNA扩增多核苷酸。对于这个方法,可以根据在此提供的序列设计出序列特异的引物,还可以购买或合成引物。用本领域已知的方法例如化学合成可以直接地合成其他的多核苷酸。
编码序列或互补序列的特别优选的部分是那些设计用作检测样品中的尼帕病毒或其他汉尼巴病毒例如亨德拉病毒的引物的部分。可以用多种报告基团或可检测部分例如放射性核素和酶标记引物,引物是那些包括尼帕病毒多核苷酸(例如SEQ ID NO:8和17)或它们的互补物的至少15、20、25、或30个连续核苷酸的引物,如在此所述的合适的引物,例如在SEQ ID NO:1中所示的序列。PCR的引物是那些包括尼帕病毒多核苷酸或它们的互补物的至少15、20、25、或25个连续核苷酸的引物,如在此所述的合适的引物,例如在SEQ ID NO:2和3中所示的序列。在一个优选的实施方式中,用于逆转录和随后扩增的引物特异地靶向于尼帕病毒基因组RNA的核壳区。
不同尼帕病毒分离株的多核苷酸和多肽序列是已知的,以及尼帕病毒的组分包括一个核壳(NC)、一个基质、一个聚合酶、一种附着糖蛋白、和P/V/C融合蛋白。这些核苷酸的实例包括那些从GenBank得到的编码为AJ564622、AJ564621、AF376747、AF212302、AY029768、和AY029767的多核苷酸。另外,在序列列表中所示的序列例如SEQ IDNO:8和17也对应于尼帕病毒多核苷酸。
同样,已经描述了尼帕病毒多肽的氨基酸序列,例如见GenBank条目AJ564622、AJ564621、AF376747、AF212302、AY029768、和AY029767。此外,特异性病毒组分的非限制性实例包括聚合酶(SEQ IDNO:9、18、28、和30);附着蛋白(SEQ ID NO:10);融合蛋白(F)(SEQ ID NO:11和20);基质蛋白(SEQ ID NO:12、21、和27);C蛋白(SEQ ID NO:13);V蛋白(SEQ ID NO:14、25和26);磷蛋白(SEQ IDNO:15、22、和24);和核壳(SEQ ID NO:16、23、31和32);糖蛋白(SEQ ID NO:19和29)。
各种亨德拉病毒分离株的多核苷酸和多肽序列以及亨德拉病毒组分都是已知的。这些多核苷酸的实例包括那些从GenBank中获得的编码为AF017149和AF 010304的多核苷酸。此外,在序列列表中所示的那些序列如SEQ ID NO:33和45也对应于亨德拉病毒多核苷酸。
同样,已经描述了亨德拉病毒多肽的氨基酸序列,例如见GenBank条目AF017149和AF 010304。此外,特异性病毒组分的非限制性实例包括核壳(SEQ ID NO:34);磷蛋白(SEQ ID NO:35和42);非结构蛋白V(SEQ ID NO:36和43);非结构蛋白C(SEQ ID NO:37和44);基质蛋白(SEQ ID NO:38);融合蛋白(SEQ ID NO:39);糖蛋白(SEQ IDNO:40);和聚合酶(SEQ ID NO:41)。
在一个实施方式中,在本发明中可以采用与在此所述的尼帕病毒或亨德拉病毒氨基酸序列具有至少70%,优选地至少80%,更优选地至少90%相同性的蛋白。
在另一个实施方式中,可以使用的尼帕病毒或亨德拉病毒蛋白是那些与尼帕病毒或亨德拉病毒编码序列有着至少70%,优选地80%,更优选地90%、95%、和97%相同性的多核苷酸序列编码的蛋白;这些多核苷酸在严格条件下将与在此所述的尼帕病毒多核苷酸序列的编码多核苷酸序列杂交。术语“严格条件”或“严格杂交条件”包括指的是在该条件下,多核苷酸将与其靶序列以比其他序列可检测到的更高的程度(例如,比背景至少高2倍)杂交。严格条件通常是那些盐浓度小于约1.5MNa离子,通常为约0.01到1.0M Na离子浓度(或其他盐)(pH为7.0到8.3)和温度至少是约30℃(短探针,例如10到50个核苷酸)以及至少是约60℃(长探针,例如超过50个核苷酸)的条件。例如,高度严格条件包括在50%甲酰胺、1M NaCl、1%SDS在37℃下杂交,以及用0.1XSSC在60到65℃下洗涤(见,Tijssen,Laboratory Techniques inBiochemistry and Molecular Biology-Hybridization with Nucleic AcidProbes,Part I,Chapter 2“Overview of principles of hybridization and thestrategy of nucleic acid probe assays”,Elsevier,New York(1993);和Current Protocols in Molecular Biology,Chapter 2,Ausubel,et al.,Eds.,Greene Publishing and Wiley-Interscience,New York(1995))。利用ClustalW算法MEGALIGNTM(Lasergene,Wisconsin)可以确定氨基酸和多核苷酸相同性、同源性和/或相似性。
更具体地,所述的检测样品中的尼帕病毒的方法包括:
用至少一种特异于所述尼帕病毒的至少一种RNA分子的引物生成所述RNA分子的DNA拷贝;
用至少一对特异于所述尼帕病毒的RNA分子的所述DNA拷贝的寡核苷酸引物扩增所述DNA拷贝;和
检测对应于尼帕病毒的扩增的DNA的存在,所述扩增的DNA的存在表示样品中存在尼帕病毒。
根据所述方法的一个有利的实施方式,所产生并扩增的DNA拷贝是尼帕病毒核壳编码区。
根据所述方法的另一个实施方式,至少其中一对寡核苷酸引物包括可检测部分。
根据所述方法的另一个实施方式,检测包括观察所述可检测部分。
根据所述方法的另一个实施方式,所述样品取自猪。
根据所述方法的另一个实施方式,所述样品取自野生动物或家养动物。
根据所述方法的另一个实施方式,所述样品取自人。
根据所述方法的另一个实施方式,其中所述至少一种特异于所述RNA分子的引物包括与编码一种多肽的多核苷酸互补的至少15个连续核苷酸,所述多肽包括选自由SEQ ID NO:16、SEQ ID NO:23、SEQ IDNO:31和SEQ ID NO:32构成的组的氨基酸序列。
根据所述方法的另一个实施方式,其中所述至少一种特异于所述RNA分子的引物包括所述多核苷酸的至少20个连续核苷酸。
根据所述方法的另一个实施方式,其中所述至少一种特异于所述RNA分子的引物包括所述多核苷酸的至少25个连续核苷酸。
具有相同性的蛋白或那些由与在此所述的多核苷酸杂交的多核苷酸编码的蛋白优选地保留了至少20%,优选地50%,更优选地至少75%和/或最优选地至少90%的野生型尼帕病毒或亨德拉病毒蛋白活性的生物活性-生物活性的量包括25%、30%、35%、40%、45%、55%、60%、65%、70%、75%、80%、85%、95%;以及在此之间的所有数值和亚界。此外,它们也可以具有相对于野生型尼帕病毒或亨德拉病毒活性的100%或更高的生物活性-生物活性的量包括至少105%、至少110%、至少125%、至少150%、和至少200%。在汉尼巴病毒属内的病毒蛋白之间的氨基酸相似性的百分比以及特别是包膜糖蛋白之间的氨基酸相似性的百分比表明了每种蛋白诱导具有交叉反应性的和交叉保护性的反应性抗体的能力。
用本领域熟知的标准技术可以将尼帕病毒或亨德拉病毒蛋白纯化到基本上纯净,这些技术包括利用例如硫酸铵等物质的选择性沉淀、柱层析、免疫纯化法等等。见,例如R.Scopes,Protein Purification:Principlesand Practice,Springer-Verlag:New York(1982)。
本发明也包括通过动员(mounting)免疫应答治疗或防护尼帕病毒引起的疾病,也包括亨德拉病毒引起的以及汉尼巴病毒属的任何成员引起的疾病。在治疗方法中,在此所述的免疫反应组合物的施用可以是用于“预防性”或“治疗性”目的。当预防性提供时,在出现任何症状之前提供免疫反应组合物。免疫反应组合物的预防性施用作用于避免、改善、和/或减轻任何随后的感染或疾病的严重度。当治疗性提供时,可以在感染或疾病的症状开始出现的时候(或不久之后)提供免疫反应组合物。因此,可以在预期暴露于致病病原体或疾病状态之前或在引发感染或疾病之后提供本发明。
将从施用在此所述的制剂中获益的对象患者在此包括任何可以从保护性对抗病毒感染中获益的动物。在一个优选的实施方式中,对象患者是人患者、马、或猪,它们是扩增宿主或经济相关动物。
可以将病毒多肽预防性地用作疫苗。本发明的疫苗含有作为活性成分的致免疫有效量的结合或融合域蛋白或在此所述的重组病毒。免疫应答可以包括生成抗体,激活抗呈现来源于病毒多肽的肽的细胞的细胞毒T淋巴细胞(CTL),或其他本领域熟知的其他机制。见例如纽约Raven出版社出版的Paul Fundamental Immunology第二版中的对免疫应答的描述(在此通过引用将其并入本申请)。有用的载体在本领域是熟知的,其包括例如甲状腺球蛋白、白蛋白例如人血清白蛋白、破伤风毒素、聚氨基酸例如聚(D-赖氨酸:D-谷氨酸)、流感、乙肝病毒核心蛋白、乙肝病毒重组疫苗。
可以将编码病毒多肽的DNA或RNA引入到患者体内以获得对多核苷酸编码的多肽的免疫应答。例如,在本实施方式中,可以施用在此所述的表达载体,并将其接种到对象患者体内以诱导免疫应答。
足以实现免疫保护或预防的量被定义为“致免疫有效量”。这种用途的有效量将依赖于组合物、给药方式、患者的体重和一般健康状态。
当与接种物有关时,术语“单位剂量”指的是适用为哺乳动物的单一剂量的物理分离的单位,每一单位都含有所计算出的生成所需致免疫作用的预定量的重组抗原或编码重组抗原的多核苷酸以及所需的稀释剂。本发明的接种物的新的单位剂量的规范是由重组病毒的特征以及所实现的特殊的免疫学作用决定的并依赖于此。
接种物通常被制备于可耐受的(可接受的)稀释剂例如盐水、磷酸缓冲盐水或其他生理的和/或可药用的稀释剂等等的溶液中,以形成一种水性的药物组合物。
接种的途径可以是静脉内、肌肉内、皮下、皮内等等,它造成了引发抗尼帕病毒的保护性应答。至少施用剂量一次。也可以施用随后的剂量。
在给哺乳动物(优选地是人)提供本发明的免疫原性组合物时,施用的剂量将有所不同,这依赖于这些因素例如哺乳动物的年龄、体重、性别、一般医学情况、既往病史、疾病进展、肿瘤负荷等等。
在免疫接种之后,可以用识别抗原的抗体或免疫细胞的生成评价疫苗的疗效,以及用特异性裂解活性或特异性细胞因子生成或用肿瘤退化评价。本领域熟练人员将知道评价前述参数的常用方法。
可以用免疫刺激剂或佐剂提高宿主的免疫应答,它们也可以被包含于免疫原性组合物中。已经鉴定出增强对抗原的免疫应答的佐剂。在人和兽医的疫苗中,常常将氢氧化铝和磷酸铝用作佐剂。其他外来佐剂和其他免疫调控物可以引发对抗原的免疫应答。这些物质包括与膜蛋白复合的皂甙以生成免疫刺激复合物(TSCOMS)、Pluronic聚合物与矿物油、矿物油中的死分枝杆菌、弗氏完全佐剂、细菌产物例如胞壁酰二肽(MDP)和脂多糖(LPS)、以及单磷酰脂质A(monophoryl lipid A)、QS21、和聚磷腈(polyphosphazene)。
在一个优选的实施方式中,分开使用尼帕病毒糖蛋白(G和F),以及在一个同样优选的实施方式中,在本发明的免疫原性组合物中联合使用G和F糖蛋白。在一个优选的实施方式中,免疫原性组合物是一种携带尼帕病毒蛋白的表达载体,其在接种之后表达引发免疫应答的蛋白,例如表达尼帕病毒糖蛋白的重组疫苗病毒,以及更优选的是表达尼帕病毒的G和F糖蛋白的载体。
本发明也设计出一种保护个体抗汉尼巴病毒感染的方法,其包括:
给所述个体或哺乳动物施用足以诱导所述个体体内的免疫应答的量的至少一种分离的汉尼巴病毒G和F糖蛋白。
根据所述方法的一个有利的实施方式,汉尼巴病毒是尼帕病毒或亨德拉病毒。
根据所述方法的另一个实施方式,所述施用还包括施用佐剂。
根据所述方法的另一个实施方式,所述施用被进行一次或多次。
根据所述方法的另一个有利的实施方式,施用至少两种汉尼巴病毒F和G糖蛋白。
本发明也涉及足以诱导免疫应答的量的至少一种分离的汉尼巴病毒G和F糖蛋白在制备用于包含个体或哺乳动物对抗汉尼巴病毒感染的药物中的用途。
根据所述用途的一个有利的实施方式,汉尼巴病毒是尼帕病毒或亨德拉病毒。
根据所述用途的另一个实施方式,所述汉尼巴病毒G和F糖蛋白与佐剂结合。
本发明也涉及一种防护或保护所需防护或保护的个体或哺乳动物对抗汉尼巴病毒感染的方法,其包括:
给所述个体或哺乳动物施用足以诱导所述个体或哺乳动物体内的免疫应答的量的表达载体(表达至少一种分离的汉尼巴病毒G和F糖蛋白),以便防护或保护个体或哺乳动物对抗汉尼巴病毒感染。
根据所述方法的一个有利的实施方式,表达载体表达至少一种汉尼巴病毒的F和G糖蛋白。
根据所述方法的另一个实施方式,表达载体是一种病毒载体。
根据所述方法的另一个实施方式,病毒载体是一种重组痘病毒载体。
根据所述方法的另一个实施方式,它还包括施用至少一种佐剂。
本发明也涉及一种表达足以诱导免疫应答的量的至少一种分离的汉尼巴病毒G和F糖蛋白的表达载体在制备用于防护或保护个体或哺乳动物对抗汉尼巴病毒感染的药物中的用途。
根据所述用途的一个有利的实施方式,表达载体至少表达汉尼巴病毒的F和G糖蛋白。
根据所述用途的另一个实施方式,表达载体是一种病毒载体。
根据所述用途的另一个实施方式,表达载体是一种重组痘病毒载体。
根据所述用途的另一个实施方式,所述表达载体与至少一种佐剂结合。
也已经开发出了抗尼帕病毒G和F蛋白以及在体外中和尼帕病毒传染性的单克隆抗体(MAb)库。此外,确信抗尼帕病毒F蛋白的抗体能中和亨德拉病毒。因此,本发明的另一个实施方式是产抗汉尼巴病毒G和F蛋白的抗体的重组杂交瘤以及表达汉尼巴病毒G和F蛋白的疫苗载体重组体。
疫苗载体重组体和杂交瘤的非限制性实例包括在2003年9月16日保藏于法国微生物保藏中心(CNCM)(Collection Nationale de Cultures deMicroorganismes,28 rue du Docteur Roux,75724 Paris Cedex 15,法国)的保藏号为I-3086的表达尼帕病毒G蛋白的重组疫苗病毒、在2003年9月16日保藏于CNCM的保藏号为I-3085的表达尼帕病毒F蛋白的重组疫苗病毒;在2004年9月9日保藏于CNCM的保藏号为I-3293的具有抗尼帕病毒的中和活性的抗尼帕病毒G蛋白的杂交瘤N°1.7;在2004年9月9日保藏于CNCM的保藏号为I-3296的具有抗尼帕病毒的中和活性的抗尼帕病毒G蛋白的杂交瘤N°3.B10;在2004年9月9日保藏于CNCM的保藏号为I-3295的具有抗尼帕病毒的中和活性的抗尼帕病毒F蛋白的杂交瘤N°35;和在2004年9月9日保藏于CNCM的保藏号为I-3294的具有抗尼帕病毒的中和活性的抗尼帕病毒F蛋白的杂交瘤N°3。
实施例1:汉尼巴病毒的金仓鼠模型
最近在马来西亚爆发了一种随后被命名为尼帕病毒(NiV)的新的副粘病毒,它感染了数百人,造成了严重的发病率以及约40%的死亡率(Chua et al.2000.Science 288:1432-1435)。患者出现了从发热和头痛到严重急性热病性脑炎综合征的症状。尽管大多数在急性感染中存活下来的有症状患者最终都康复,且没有严重的后遗症,但是仍有少数患者在数月和数年后因复发性脑炎而再次入院(Tan et al.Ann Neurol.2002.51:703-708)。发现复发性脑炎的临床特点和发病机理都不同于急性NiV脑炎。现在已经很好地建立了经紧密接触而造成的猪-人传播,猪扮演着病毒的部分扩增宿主的作用(Parashar et al.J Infect Dis.2000.181:1755-1759)。天然宿主很可能是果蝠,因为最近从果蝠的尿液中分离出了NiV(Chuaet al.Microbes Infect.2002.4:145-151)。因此,NiV爆发代表着最为严重的最近已经出现于蝠类的病毒性动物传染病(Eaton,Microbes Infect.2001.3:277-278)。
根据对NiV感染的人体组织的研究,开始了解了NiV感染的病理和致病机理(Wong et al.Am J Pathol.2002.61:2153-2167)。在急性NiV感染中,现有的证据说明在初期病毒复制之后,出现了病毒血症,病毒随之在全身播散。血管被感染,造成了广泛的血管炎,这导致了多个器官中的血栓形成、血管阻塞、缺血和/或微坏死,影响中枢神经系统(CNS)最为严重(Wong et al.Am J Pathol.2002.61:2153-2167)。血管外的实质组织(最主要是神经元)也对感染易感。已经假定CNS缺血和/或微梗死、和直接的神经元感染的组合可以造成在急性NiV感染中所见的严重的神经表现(Wong et al.Am J Pathol.2002.61:2153-2167)。
动物模型的缺乏阻碍了对进一步了解急性NiV感染的早期发病机理的尝试。现有的急性NiV感染的病理及发病机理的知识都与疾病晚期有关,因为研究都依据于人体尸解。至今已经被研究的天然和试验感染的动物包括猪和猫都表现为血管炎,但没有表现出在人NiV感染中所发现的典型的脑炎,因此它们可能不适合作为动物模型(Hooper et al.2001.Microbes Infect.3:315-322)。用作感染患者中的经验治疗并被报道为有效的抗病毒的利巴韦林还必须在动物试验中进行充分的评估(Chong et al.,Ann Neurol.2001.49:810-813)。同样,因为缺乏好的模型,也不能测试其他的抗病毒药物和新开发的疫苗在NiV感染中的潜在用处。可以在动物模型中进行可控制的传染研究,以研究病毒传染性和传染途径。
在这个研究中,我们研究了一些作为急性NiV感染的潜在模型的动物物种,并将金仓鼠(Mesocricetus auratus)鉴定为一种合适的模型。用各种方法特征性地描述了经鼻内和腹腔内感染的金仓鼠的病理损害,其病理损害表现出与在人体疾病中所发现的病理改变的高度相似性。我们也尝试联系在各种具有在此所发现的病理改变的感染器官中的病毒分离和病毒基因组检测。
材料和方法
病毒原液和滴定
在Vero细胞中2次传代之后,法国里昂的BSL-4“Jean Mérieux”实验室收到了KB Chua博士和SK Lam博士的从患者的脑脊液中分离到的NiV。在4级物理防范下进行Vero细胞中的第3次传代之后得到了病毒原液。
在感染后1-2天,当Vero细胞表现出融合及合胞体形成时,收集病毒的上清液。通过将每个孔内的200μl上清液的系列的10倍稀释液在37℃下孵育1个小时,在6孔板中滴定病毒原液。用Dulbecco最低基本培养基(DMEM)洗涤每孔细胞两次,并往每孔内加入2ml含有2%胎牛血清的1.6%羧甲基纤维素DMEM溶液。将板在37℃下孵育5个小时,用磷酸缓冲液(PBS)(pH 7.4)洗涤孔,并用10%福尔马林固定20分钟,洗涤并用亚甲兰染色。在用0.01的感染复数(MOI)感染24个小时之后,上清液中的病毒滴度是2×107噬斑形成单位(pfu)/ml。
动物感染试验
一起进行3个系列的动物研究。在第一个研究中,在两组动物中进行对NiV感染的易感性的预试验,两组动物每组都包括5只小鼠、2只豚鼠和2只仓鼠。在这个试验中使用了4周龄的、雌性Swiss小鼠(Charles River,L’Arbresle,France),4个月大的、雄性Hartley豚鼠(Charles River)、和2个月的雄性金仓鼠(Janvier,Le Fenest St Isle,France),其中经鼻内(IN)或经腹腔内(IP)接种每组动物。对于IN途径,每只动物给予30μl病毒原液(6×105pfu),而对于IP途径,接种0.5ml(107pfu)。观察动物的感染征象。将动物圈养在BSL-4实验室的动物房的装配有Hepa滤器的通风防范笼内。我们遵循着法国的处理动物条例,以及在高度安全的BSL-4防范区内工作所要求的强制规范。
根据第一个研究的结果,然后利用IN和IP接种途径在成体仓鼠(7-14周龄)中进行第二个研究,以确定杀死50%动物所必需的致死剂量(LD50)。用NiV原液的10倍稀释液感染6只仓鼠的组,并每天观察两次共4周。
为了研究圈养在同一笼内的动物之间的持续的再感染造成死亡率的可能性,进行了第三个研究。在这个研究中,用105pfu病毒经IP途径感染2只仓鼠,并将其在接种后与4只其他未感染的仓鼠放置在同一笼内3天。观察动物,并在30天后取眶后窦血样进行血清学检查。
从总共12只最近死亡的(≤12hours)或终末濒死的仓鼠中收集第一个和第二个研究中的合适的组织样品包括血液、脑、肺、心脏、肝脏、脊髓、脾和肾脏。用氯胺酮和甲苯噻嗪麻醉终末濒死仓鼠,用心脏穿刺放血并尸解。只要有可能就从膀胱收集尿液。不研究发现在超过12个小时后死亡的动物。
将组织冷冻在-80℃,用于病毒培养和逆转录聚合酶链反应(RT-PCR)分析。对于组织病理学研究,在BSL-4实验室外进行常规组织处理和石蜡包埋之前,先将组织在10%缓冲福尔马林中固定至少15天。也从福尔马林固定的尸体中切下鼻道和颈部淋巴结的组织,将其只用于常规处理和石蜡包埋。对于电镜(EM),将新鲜的或福尔马林固定的组织在3%戊二醛(0.1M磷酸缓冲液,pH 7.4)固定数个小时,再转移到磷酸缓冲液中。同样,将用于免疫电镜(IEM)的组织在2%多聚甲醛/0.05%戊二醛中固定,再转移到缓冲液中。另外,用伽马照射(2×106拉德)最初没有被福尔马林固定的EM和IEM组织以进一步保证没有传染性。
在尸解时通过心脏穿刺收集血样,或在第二个研究中,在感染后4周从存活动物的眶后窦中取血。根据Reed和Muench方法计算出引起50%仓鼠死亡率的NiV剂量。
病毒分离和滴定
用Vero细胞的噬斑滴定测定尿液和其他组织中的感染病毒颗粒的量。在2ml管中机械粉碎(Mini-beadbeater;Biospec,Bartlesville,USA)每种器官的一小片段两次,每次30秒,每2ml管含有0.5ml无菌玻璃珠和0.5ml DMEM。在4℃下,将管在3000rpm离心5分钟,并将200μl上清液的系列稀释液铺板在6孔板的用于病毒滴定的Vero细胞上。
尼帕病毒抗体测定
用酶联免疫吸附检测法(ELISA)个别地测定感染仓鼠的血清中是否存在NiV抗体。从用0.01pfu/细胞的MOI感染24个小时的感染Vero细胞中制备出NiV抗原的粗提取物。用PBS洗涤细胞,并在4℃将其在含有1%Triton X100的PBS中裂解10分钟(107细胞/ml)。将细胞裂解液超声处理2次,每次30秒,以充分地破碎细胞,并将其在4℃、5000rpm离心10分钟。将上清液冻存在-80℃。相似地处理未感染的Vero细胞以制备抗原对照。用来自恢复期的NiV感染患者的血清进行对尼帕病毒抗原的交叉滴定,以确定对应于表现为最高O.D.读数的稀释液的抗原滴度。
逆转录聚合酶链反应
用RNA提取试剂盒(QIAamp Viral RNA Mini Kit;Qiagen Inc.,Valencia,California,USA)从20μl血清和尿液、以及从机械粉碎的、新鲜冷冻的组织中提取出总RNA。在RT-PCR(Titan One Tube RT-PCRSystem;Roche Diagnostics,Mannheim,Germany)操作中施用约2μg提取物,以检测NiV核蛋白(N)基因的存在。特异引物以前已经发表(Chuaet al.Science.2000.288:1432-1435)。
光镜
将福尔马林固定的、石蜡包埋的组织切成3μm厚的切片,放置于玻璃载玻片上,并用hemalin-phloxine-safranin染料染色,用于光镜检查。
免疫组织化学(IHC)
将3μm厚的组织切片放置在硅烷载玻片上,并用二甲苯和分级(graded)乙醇洗涤剂脱蜡。用热处理(pH 6.0的柠檬酸缓冲液,96-98℃处理40分钟)复原抗原。在冷却到室温(20℃)之后,始终在20℃,如下连续地孵育切片,在各个步骤之间用PBS洗涤:(a)4%牛血清白蛋白/10%山羊血清(GS)PBS溶液,15分钟;(b)兔源的、多克隆抗NiV抗体,1∶500,1个小时;(c)生物素化的、山羊抗兔二抗,30分钟(Dako,Trappes,France);(d)0.09%H2O2 PBS溶液;(e)辣根过氧化物酶连接的抗生物素蛋白链菌素和二氨基联苯底物(按照产品说明书,Dako,Trappes,France)。将载玻片在苏木精中复染,并将其包埋在水性介质中(Aquamount,Merck Eurolab,Strasbourg,France)。
原位杂交
对于ISH,从228个碱基对的、利用尼帕病毒特异性引物的RT-PCR产物(Chua et al.Science.2000.288:1432-1435)生成地高辛(DIG)标记的放射性探针。根据产品说明,将该片段克隆于pdrive克隆载体中(Qiagen PCR cloning kit,Qiagen Inc.,Valencia,California,USA)。用限制性内切酶Hind III将含有两个方向上的正确插入物的质粒线性化,并将其转录,以便用DIG RNA标记试剂盒(Roche Diagnostics,Mannheim,Germany)生成有义的和反义的放射性探针。用DNase处理放射性探针(15分钟,37℃),然后在使用之前用乙醇沉淀法纯化。
用0.2N HCl(20分钟,20℃)预处理脱蜡的组织切片,随后用0.1mg/ml蛋白酶K的100mM Tris/50mM EDTA缓冲液(pH 8.0)(15分钟,37℃)处理。在2次PBS洗涤之后,在45℃保湿皿(HybaidOmnislide)内将载玻片与经过滤的杂交溶液中的放射性探针的1∶50到1∶100稀释液孵育过夜,杂交溶液含有45%甲酰胺、6xSSC(1xSSC=0.15M氯化钠,0.015M柠檬酸钠,pH 7.0)、5x Denhardt溶液、100μg/ml变性的鲑鱼精子、100μg/ml酵母菌tRNA和10%硫酸葡聚糖。
序列的杂交后步骤包括(a)6xSSC(3x20分钟,45℃);(b)2xSSC(10分钟,20℃);(c)100mM Tris,pH 7.5/150mM NaCl缓冲液(1分钟,20℃);(d)相同的Tris/NaCl缓冲液和2%GS及0.1%Triton(30分钟,20℃)。然后将载玻片与1∶1000稀释于Tris/NaCl/GS/Triton缓冲液的碱性磷酸酶缀合的、抗DIG Fab片段(Roche diagnostics,Mannheim,Germany)在保湿皿中孵育(过夜,20℃)。按照产品说明,在与含有NBT/BCIP溶液(Roche Diagnostics,Mannheim,Germany)的Tris/NaCl/MgCl2缓冲液一起孵育之前,用Tris/NaCl(pH 7.5)缓冲液(3x10分钟)和100mM Tris(pH 9.0)/150mM NaCl/50mM MgCl2缓冲液(1分钟)洗涤终止反应。在约45分钟之后,用10mM Tris(pH 8.0)终止显色反应。用苏木精复染载玻片,并在水性介质中用盖玻片将其盖上。
动物感染试验:存活和LD50
在第一个研究中,没有一只经IN或IP途径接种的Swiss小鼠出现任何临床征象。只有经IP途径感染并因此接受了107个感染性病毒颗粒的Hartley豚鼠在5-7天后表现出短暂的发热和体重下降,但是它们可康复。经两种途径感染的金仓鼠都表现出运动和平衡困难,并在感染后5-8天快速死亡。
图1显示了在第二个研究中的金仓鼠的剂量-生存图,用病毒的系列稀释液(即经IP途径:1到104pfu和经IN途径:10到106pfu)接种金仓鼠。经IP感染的金仓鼠的感染与出现临床征象及死亡之间的时间间隔更短,它们在感染后5到9天以及在出现震颤及肢体麻痹后<24h内死亡。相反地,绝大多数IN接种的动物表现出进行性恶化,表现为平衡失调、肢体麻痹、昏睡、肌肉颤搐和终末期的呼吸困难。绝大多数动物在9到15天之间死亡。但是,6只动物死得更晚,1只死于第18天、2只死于第21天以及3只死于第29天。IP和IN途径的动物的LD50分别是每只动物270pfu和47,000pfu。
在那些用更小剂量(IP途径1和10pfu/动物;IN途径10 and 102pfu/动物)接种的、在感染后存活超过30天的动物中,没有出现血清转换现象(数据没有显示)。事实上,这些动物没有一只死亡或表现出任何疾病的征象。相反地,用更高病毒剂量感染的与被给予相同剂量并死亡的动物饲养在同一笼子中的存活动物已经具有了高水平的抗体(数据没有显示)。但是,这些幸存者都没有表现出疾病的临床征象。
在传染研究(第三个研究)中,其中未感染的动物与感染的动物饲养在一起,没有一只未感染的动物表现出疾病或血清转换的证据(数据没有显示)。
病毒分离和病毒基因组检测
一般而言,对在尸解中取得的各种动物标本的RT-PCR显示在绝大多数组织和尿液中都能检测到NiV病毒基因组(表2)。血清是特别例外的,其中血清中的病毒基因组都为阴性。因此,没有在血清中尝试进行病毒培养。在进行这两种测试的组织中,病毒培养阳性的组织的范围与RT-PCR有着很好的相关性,尽管病毒培养的阳性百分比更低,特别是在经鼻内感染的仓鼠中。
病理特点
血管
在多个器官包括脑、肺、肝、肾脏和心脏研究了血管病理。在大血管中,更显著变化的特点是局灶的、透壁的纤维素样坏死以及周围的炎症(图2A)。但是,血管炎可能是更细微的,具有更少的炎症细胞(图2E、3A)以及非常局灶的核固缩和核碎裂(图2E)。在一只在经腹腔内接种后8天死亡的仓鼠中,看到了内皮生成的多核合胞体(图2C)。在一些血管的血管腔内可以发现血栓形成(图4B)。如IHC和ISH分别证实的那样,病毒抗原和基因组定位于内皮细胞和合胞体,以及下面的血管中膜的平滑肌(图2D、F)。在血管壁上检测到了病毒核壳。
中枢神经系统
与其他器官比较,脑是在血管和实质损害方面影响最为严重的器官。除血管炎之外,最显著的征象是出现在通常在血管炎附近的神经元上。受累神经元表现为细胞浆内的多个嗜酸性包涵体(图3C)。这些包涵体以及没有显著包涵体的神经元胞浆和神经元突出常常是病毒抗原和RNA均为阳性(图3D-F)。在超结构上,这些包涵体是由确定量的通常与副粘病毒相关的细毛(fuzzy)型的丝状核壳构成(图5A)。用NiV特异性抗体免疫标记这些包涵体(图5B)。没有发现核包涵体,但是有着核IHC阳性的证据(图3D,插图)。
其他的实质改变包括具有缺血/梗死和水肿证据的局灶区域(图3B)。实质和脑膜的炎症通常是轻微的,仅仅偶尔观察到了血管周围白细胞套袖现象和噬神经细胞作用。很罕见地,在室管膜内层(图3E)、以及在脑膜及脉络丛中发现的单个核细胞中观察到了IHC阳性。但是,脉络丛被覆上皮的病毒抗原和基因组均为阴性。在白质中没有观察到IHC和ISH阳性。
其他器官
在肺中,有时可以观察到通常与血管炎的血管相关的实质炎症的小的单个结节或更为融合的区域(图4A、B)。炎症细胞主要由巨噬细胞、中性粒细胞和淋巴细胞的不同的混和物构成。经IHC和ISH证实的NiV阳性的多核巨细胞核炎症细胞是罕见的。肺实质的纤维素样坏死并不明显。没有发现支气管炎、多核合胞体或NiV感染支气管上皮的其他证据。
肾脏的肾小球损害是罕见的,但是最显著的损害是肾小球毛细血管中的血栓性栓子、外周的多核合胞体、以及周围的炎症(图4C)。仅仅在少数肾小球和小管中检测到病毒抗原(图4D)。在一些动物的肾脏中,伸入到肾盂内的肾乳头的覆盖上皮始终都证实了病毒抗原的存在(图4E),但是在同一上皮上却没有ISH。
注意到了脾脏上的罕见的坏死病灶,但是没有观察到血管炎或多核巨细胞。IHC和ISH在小动脉周围的淋巴细胞上偶尔阳性(图4F)。没有出现特异的肝实质损害。在心脏中,仅仅罕见地观察到了与梗死相关的心肌炎(图2B)。在淋巴结或鼻上皮中都没有检测到炎症或病毒抗原。
在三种用NiV接种的动物物种(即小鼠、豚鼠和仓鼠)中,仓鼠表现得最为易感。根据途径和剂量,大多数感染仓鼠都发生了严重的病变。对从感染仓鼠中得到的组织的研究说明它是一种急性NiV感染的合适的动物模型,它证实具有大多数在人急性NiV感染中发现的特征。
可以经IP或IN途径感染仓鼠,但是经IP途径的感染比经IN途径杀死动物更快。此外,需要远为更低的IP剂量杀死相同数目的动物,如IP和IN感染动物之间的十分不同的LD50剂量所显示的那样。这可能不是奇怪的,因为IN接种的NiV在发生感染之前可能必须要穿透气道-消化道的粘膜屏障,而IP接种的NiV理论上可以直接进入全身循环。
对感染仓鼠组织的组织病理学研究显示血管(特别是那些CNS血管)发生特征为坏死和壁内炎症的血管炎。通过内皮的多核包涵体形成的存在以及对血管壁内的病毒核壳体、抗原和基因组的检测提供了病毒直接感染血管壁(包括内皮和平滑肌)的证据。更可能是血管炎的结果是,所发生的血栓形成及血管阻塞造成了脑和心脏的远端缺血和微梗死。尽管程度更轻,但是血管炎也累及肺和肾脏的血管,以及梗死是不明显的。这些发现与在人感染中所见的相似(Wong et al;Am J Path.2002.161:2153-2167),明显的例外可以是在人感染中没有报告有肝脏的血管炎。
除了缺血和梗死,CNS神经元也表现出感染的证据,如存在神经元的病毒包涵体、抗原和基因组。主要在细胞浆内发现的病毒包涵体是由典型的副粘病毒型核壳构成。在CNS的血管、实质和神经元中的发现使得它们成为急性NiV感染的主要靶点,患病动物具有显著的CNS征象例如麻痹、步态和平衡异常的事实也支持这一点。对于人感染,CNS症状和征象是非常明显的,并且CNS也是受累最为严重的器官(Gooh et al.N Engl J Med.2000.342:1229-1235;Wong et al;Am J Path.2002.161:2153-2167)。
在仓鼠肾脏中,血管炎和肾小球损害与在人患者中报告的损害相似(Chua et al.Lancet 1999.354:1257-1259;Wong et al;Am J Path.2002.161:2153-2167)。病毒抗原而不是病毒基因组在肾乳头的覆盖上皮中的始终存在说明可能是重吸收了漏到尿液中的IHC可检测的病毒蛋白。Williamson等发现了在亨德拉病毒感染的豚鼠膀胱中的泌尿道感染的证据,但是没有关于肾脏中的上皮感染的信息。病毒抗原和基因组在脾小动脉周围淋巴细胞中的出现说明在此处发生了活动的病毒复制。在仓鼠心脏中,推测罕见的梗死可能与和人相似的血管炎有关(Wong et al;AmJ Path.2002.161:2153-2167)。
关于NiV感染动物的有限的发表数据(包括对野地和试验感染的猪和猫、以及野地感人那的狗和马的观察)显示系统性血管炎是所有这些动物中的常见特征(Hooper et al.Microbes Infect.2001.3:315-322)。但是,它显示这些动物中没有一只具有如我们的研究在仓鼠上所令人信服地证实的脑炎及神经元感染。对于猪和猫,虽然有着脑膜炎的证据,但是没有显著的脑炎或任何神经元感染的明显的直接证据。在狗和马中,除了脑膜炎,也发现了局灶的脑实质疏松现象,但是并没有任何关于脑炎或直接神经元感染存在的数据。因此,这些动物都不是急性人类疾病的好模型,它的典型表现是显著的血管炎脑炎和直接神经元感染。
通过在所有测试的实体器官中的病毒分离和/或RT-PCR确认了经IHC和ISH的病毒组织定位。总而言之,作为一种用于IN和IP感染动物中的NiV感染的确认试验,RT-PCR比病毒分离更为敏感。从IN感染动物中的病毒分离率比IP感染动物更低,可能是因为前者的存活时间更长,它可能有利于对实体器官中的病毒的有效的免疫清除。但是,不论存活时间有多长,在所有测试的7只动物的血清中的RT-PCR均为阴性,这说明免疫系统清除循环中的病毒或在感染早期发生的病毒血症可能是更为有效的。同样,病毒颗粒可以被转运到感染的血白细胞内。需要仓鼠模型中的其他研究来澄清这一点。
在以前的人体研究中,根据多个器官和弥漫血管的同时受累,以及对血管损害例如血管炎、血栓形成和梗死发生得比血管外实质损害更早的观察也推测早期已经发生了病毒血症(Wong et al;Am J Path.2002.161:2153-2167)。我们的数据确证了这些发现,它也显示出同步的及广泛的器官受累。
RT-PCR和病毒分离所确认的尿液中的病毒的存在与肾脏肾小球损伤很好地相关联。从人患者中已经报告了人体尿液中的病毒排泄物,并推测这是病毒传染给医务工作者的可能方式。经口消化和/或喷雾吸入被感染的分泌物被认为是造成了猪-人的病毒传染(Parashar et al.J InfectDis 2000.181:1755-1759)。经IN途径对仓鼠的成功感染支持这一点。
一种急性NiV感染的动物模型的建立应当打通了更好地了解其发病机理特别是关于早期事件的发病机理的道路,因为目前的NiV知识主要依据于晚期病变。也可以在模型中测试潜在的抗NiV药物和疫苗的疗效。对免疫应答的更好的了解能让我们研究是否NiV能引起免疫抑制,这是一种在麻疹感染中熟知的现象。用于复发性NiV脑炎的动物模型仍是难以捉摸的,但是对最终康复的大量感染仓鼠的长期随诊能生成一些复发性脑炎的病例,因为人复发性脑炎的发生率是低的(Tan et al.AnnNeurol.2002.51:703-708)。
实施例2:利用实时PCR对汉尼巴病毒RNA的特异的及敏感的定量检测
尼帕病毒被分级为第4级病原体,所有的试验都在里昂的4级生物安全水平(BSL-4)的Jean Merieux实验室进行。根据生物安全规范,只在BSL4实验室外测试RNA提取物。
细胞和病毒
尼帕病毒(从患者的脑脊液中分离到的)是Kaw Bing Chua博士和Sai Kit Lam博士(Kuala Lumpur,马来西亚)的馈赠。通过在BSL-4实验室中用感染复数(MOI)为0.01噬斑形成单位(pfu)/细胞感染Vero-E6细胞制备病毒原液,并在感染后24小时回收病毒。病毒滴度为2×107pfu/ml。
在用0.01MOI的尼帕病毒感染的Vero细胞中监测病毒生成的时相。Lab-tek培养板中的亚融合细胞的孔被尼帕病毒感染或模拟感染。在37℃孵育1小时之后,用Dulbeco最低基本培养基(DMEM)洗涤细胞三次,并往每孔内加入0.5ml含有2%胎牛血清(FCS)的DMEM。在4天内每天收集每孔中的上清液并将其转移到Eppendorf管内,在2000rpm离心5分钟,然后分装到两个新管内。将一组含有感染的或模拟感染的细胞的上清液的管处理用于RNA提取及定量,而另一组管被用于病毒滴定。
将每孔内的单层细胞在10%福尔马林中固定20分钟,以及在0.1%Triton X100中固定5分钟。用PBS冲洗细胞,并将其在37℃下与含有抗尼帕病毒抗体的人康复期血清的稀释液孵育30分钟。然后冲洗细胞,并将其与含有0.1‰碘化丙啶溶液的荧光素缀合的抗人IgG抗体孵育。在末次冲洗之后,在UV显微镜(Leica)下观察细胞。
动物
用5×104pfu(约100×LD50)(Wong,et al 2003.Am.J.Pathol.)经腹腔内感染5只7到14周龄的金仓鼠(Janvier,Le Fenest St Isles,France)。在感染后第5天时,经眼穿刺取得每只动物的血样,并将血清储存在-80℃直到使用。我们遵循法国的处置动物的规定,以及在BSL-4防护内工作的强制规范。
病毒滴定
用Vero细胞噬斑检测滴定病毒。简单地,将含有亚融合的Vero细胞的6孔板在37℃、5%CO2孵箱内与1ml病毒原液的系列稀释液(用1∶10作为起始稀释,对于仓鼠血清为1∶100)孵育1个小时。用没有FCS的DMEM洗涤细胞两次,并用2ml 1.6%羧甲基纤维素的含5%FCS的DMEM覆盖细胞。在37℃孵育5天后,用10%福尔马林固定细胞,用亚甲兰染色并用水冲洗细胞。计数噬斑并将滴度表示为pfu/ml。
RNA提取
根据产品说明,用RNA提取试剂盒(QIAamp Viral RNA Mini Kit,Qiagen Inc.,Valencia CA,USA)从140μl尼帕病毒感染的Vero细胞的上清液或从20μl仓鼠血清中提取病毒RNA。将提取物重悬在60μl AVE缓冲液中,分装并储存在-80℃,直到进行RT-PCR扩增。
阳性尼帕病毒对照的制备
将整个尼帕病毒NP基因克隆到含有T7启动子的PCR TA克隆载体pDrive(Qiagen)中。用DNA测序(Big Dye Terminator,AppliedBiosystems,USA)确认插入物的序列和位置。在利用T7 RNA聚合酶(Invitrogen)体外转录之前,在NP基因的末端将质粒pDrive-NP-NiV线性化,然后用Geneclean_II试剂盒(Q-Biogene)纯化。用无RNase的DNase I(Roche diagnostics)处理RNA转录产物以去除DNA模板,然后用RNA NOW(Ozyme)提取,并用乙醇沉淀。将RNA重悬在水中,并储存于-80℃。为了确保模板DNA已经被清除,在用无RNase的DNaseI处理之前和之后,用TaqManTM PCR系统(TaqManTM universal PCRMaster Mix 200RXN,Applied Biosystems)进行定量PCR检测。用分光光度计确定RNA的量并用所测定的量实现对实时RNA定量的标准曲线。
引物和TaqManTM探针
按照所推荐的标准,用程序Primer ExpressTM(Perkin-Elmer,AppliedBiosystems,USA)设计出尼帕病毒NP基因的引物和探针。前向引物(Ni-NP1209 5’GCAAGAGAGTAATGTTCAGGCTAGAG 3’-SEQ ID NO:1)和逆向引物(Ni-NP1314 5’CTGTTCTATAGGTTCTTCCCCTTCAT 3’-SEQID NO:2)扩增出了一个105个碱基对片段。设计出荧光探针(Ni-NP 1248Fam 5’TGCAGGAGGTGTGCTCATTGGAGG 3’-SEQ ID NO:3),其与在PCR引物内侧的序列退火。荧光报告染料6-羧基-荧光(FAM)位于探针的5’末端,以及猝灭剂6-羧基-四甲基-罗丹明(TAMRA)位于3’末端。
RT-PCR TaqManTM反应
利用ABI PRISM 7700 TaqManTM系列检测仪进行定量RT-PCR检测。用一步RT-PCR系统(TaqManTM一步PCR基本混和试剂试剂盒,Applied Biosystems)进行一个连续的热循环。制备基本(master)混和反应物并将20μl部分或22.5μl部分分装到薄壁的microAmp光度管(ABIPRISMTM,Applied Biosystems)中。往每个管内加入5μl仓鼠血清的RNA提取物、或2.5μl原液病毒或感染细胞上清液的RNA提取物、或2.5μlRNA转录产物。最终反应混和物含有900nM每种引物和200nM每种探针。在扩增之前,将RNA在50℃下逆转录30分钟。接着是一个循环的94℃下变性5分钟,接下来,进行45个周期的94℃下15秒和60℃下1分钟的PCR扩增。在第二个步骤结束时,读取荧光,这容许连续地检测RNA的量。阈值循环(Ct)是在发射的荧光超过所谓的“检测阈值”(Dt)的固定界限之前的循环的数目。Dt的确定是依据于其中能检测病毒RNA并仍在标准曲线的直线范围之内的最低水平。因此,最初存在的靶点数目的log10与Ct值成正比,并可以用标准曲线将其测定。
将来自麻疹病毒株CR68的RNA(其性质已经被验证)用作阴性对照。
这些试验显示了一种用于检测并定量尼帕病毒RNA的检测法,它是多能的、高度可重复的并且随着时间是稳定的。为了实现这一点,我们已经开发出了一种尼帕病毒TaqManTM RT-PCR检测法。
检测法的敏感性和特异性
用一系列含有从尼帕病毒原液中提取的RNA的稀释液的样品评价了尼帕病毒检测法的敏感性和特异性。测试了一组含有从1.2×105pfu到0.12pfu/管(体积为2.5μl)的10倍病毒稀释液。从这组稀释液的扩增曲线中计算出阈值循环(Ct)值(图5)。图6显示检测是线性的,从1.2×105pfu到1.2pfu/次。这说明扩增试验对大范围的病毒滴度的可行性以及其敏感性。当重复三次试验时,得到了相似的数据,说明了检测的可重复性(数据没有显示)。利用麻疹病毒RNA和尼帕病毒引物及探针没有发生扩增也验证了检测的特异性(数据没有显示)。
为了将检测法标准化,用RT-PCR TaqManTM测试了已知量的在体外从质粒pDrive-NP-NiV转录的RNA的系列稀释物。用三种利用在不同日期制备的转录RNA的检测法绘制标准曲线(图7)。曲线的线性容许定量每个反应的109 to 103个RNA分子。此外,小偏差(R2=0.9834)说明检测法是高度可重复的(图7)。发现通过比较从两种RNA转录产物中获得的Ct值计算出的检测间的变异系数有所不同,在0.3到2.2%之间。
感染细胞上清液中的病毒负荷的定量
为了确定我们的用于定量尼帕病毒RNA的TaqManTM RT-PCR方法的准确性,将噬斑检测法得到的感染病毒滴度与通过利用RNA转录标准曲线的TaqManTM RT-PC计算出的基因组等价物的量进行比较。用感染复数MOI为0.01pfu/小细胞的尼帕病毒感染Vero细胞,并分析在感染后第1、2、3、和4天取得的细胞上清液。在感染后第1天已经观察到了轻微的病毒诱导的细胞病变效应,细胞融合的数目和强度每天都有所增强,直到感染后第4天细胞破坏是完全的(图8)。培养基中的感染病毒和病毒RNA的量有所增加直到各种感染的第3天,然后逐步下降(图9)。此外,感染颗粒的数目与RNA基因组的数目之间的RNA/pfu比值不是恒定的,随着感染的时间而增加(表1)。
表1:用噬斑检测法和实时RT-PCR检测法检测感染细胞上清液中的感染尼帕病毒和尼帕病毒RNA。
感染后天数a病毒RNA/ml(×10-6)b pfu/ml(×10-3)RNA/pfuc
测试次数 | 1 | 2 | 1 | 2 | 1 | 2 |
1234 | 1176019241549 | 520073353NT* | 217751210400 | 920372275NT | 53898115903872 | 50710491473NT |
a-以0.01的MOI感染细胞,并在感染后第1、2、3和4天分析上清液。
b-用RNA转录标准曲线计算出尼帕病毒RNA的浓度。
c-在Vero细胞上清液中检测到的感染颗粒的数目与病毒RNA的数目之间的RNA/pfu比值。
*没有测定
为了确认样品中的病毒RNA分子的数目的准确性,用TaqManTM分析在感染后第3天提取的RNA的10倍稀释液,并将其与pfu的理论数目进行比较(表2)。选择第3天是因为它对应于RNA和感染病毒产生的峰值。在经稀释的感染后第3天时的样品中得到的RNA/pfu比值有所增加,这与病毒RNA的量的变化相反。
表2:对来自经稀释的用0.01pfu/细胞感染后第3天的感染细胞上清液的尼帕病毒RNA的实时定量
RT-PCR TaqManTM和RT-PCR.
测试1 | 测试2 | |||||
pfu/ml(×10-3)121012112.11.21 | RNA/ml(×106)1738(1924)a229314 | RNA/pfu1436(1590)189125433349 | RT-PCR++++ | pfu/ml(×10-3)2275227.522.752.2750.22750.0228 | RNA/ml(×106)3090(3353)a30836.93.90.59UDb | RNA/pfu1358(1473)1353162217142588 |
a-根据在表1中所述的试验计算出刮弧内的值。
b-未定量的数据(在样品中检测到RNA,但是Ct值却是在标准的直线范围之外)
对经尼帕病毒感染的仓鼠的血清中的病毒RNA的检测
为了评价我们的尼帕病毒TaqManTM检测法是否容许检测并定量生物样品中的病毒RNA,用噬斑滴定和实时RT-PCR分析5只经尼帕病毒感染的仓鼠的血清。以前的研究已经显示在感染后第5天可以检测到仓鼠中的病毒血症。结果(表3)说明在三只动物中检测到病毒RNA以及在两只动物中检测到感染病毒。病毒基因组分子的数目比活病毒的数目高约3logs。
表3:对感染后第5天提取的感染仓鼠血清中的尼帕病毒RNA的检测
仓鼠 | ARN/ml(10-3) | Pfu/ml | RNA/pfu |
H1H2H3H4H5 | 7051413628NDaND | 500500NDNDND | 1410826 |
a-未检测
用杀死50%动物所需剂量的100倍剂量经腹腔内感染仓鼠。用利用RNA转录产物的曲线计算出对扩增曲线的定量。
已经开发出的检测法提供了一种对尼帕病毒感染的快速的、准确的和定量的诊断。这个测试对于需要确认临床或野地样品中的尼帕病毒病原学的实验室是一种有用的工具。尼帕病毒是一种对人高度致病的病毒,它已经杀死了超过40%的感染个体(Goh,et al 2000,New Engl J Med.342:1229-35;Chong,et al 2002,Can J Neurol Sci.29:83-7;Lee,et al 1999,Ann Neurol.46:428-32)。在猪中,死亡率是低的,但是因为感染率接近100%,为了阻止尼帕病毒的流行,1999年在马来西亚屠杀了超过100万只猪,这对国家的养猪工业造成了毁灭性的影响(Mohd Nor et al 2000,Rev Sci Tech Off Int Epiz.19(1):160-5;Chua,2000,Science.288:1432-5)。尽管自从马来西亚和新加坡的末次流行之后,还没有确定出新的人或猪的病例,但是2001年在加拿大发现了携带抗尼帕病毒抗体的翅状蝙蝠(Pteroid bats),这说明病毒可能在任何时间在东南亚再次出现。2001年在孟加拉国和印度北部报告了尼帕病毒样疾病,但是还没有得到关于致病病原体的性质的确切数据(ProMed 2002 Nipah-like virus-Bangladesh(2001):Archive number 20020830.5187;ProMed 2003 Nipah-like virus-India(North Bengal):2001 Archive number 20030106.005027)。对这种病毒的阳性鉴定对于实施合理的控制手段是必需的。但是,治疗或抗该病原体的疫苗的缺乏迫使其在细胞培养物中的繁殖(用于病毒分离和鉴定、血清中和、和ELISA的抗原制备)只能在生物安全水平BSL-4实验室中进行。这些限制将限制对人脑炎的研究以及对野生和家养动物的生物样品中的病毒检测。为了保证操作者的安全,尼帕病毒的诊断性实时PCR检测法的应用将是一种用于初步鉴定样品的首要的安全的方法,然后可以在BSL-4试验中处理样品以繁殖病毒。
已经开发出了诊断大量病毒例如水痘带状疱疹病毒、人乳头瘤病毒、丙型肝炎病毒、登革热病毒、Epstein-Barr病毒或流感病毒的TaqManTM检测法(Hawrami,et al 1999,J Virol Methods.79:33-40;Josefsson,et al 1999,J Clin Microbiol.37:490-496;Morris,et al 1996,JClin Microbiol.34:2933-2936;Laue,et al 1999,J Clin Microbiol.37:2543-2547;Leung,et al 2002,J Immu Methods.270:259-267;Schweiger,et al2000,J Clin Microbiol.38:1552-1558),并且已经用这项技术帮助诊断一些致命性的地方性传染的蚊媒的和出血性的病毒疾病(Lanciotti,et al2000,J Clin Microbiol.38:4066-4071;Garin,2001,Microbes Infect.3:739-745;Garcia,et al 2001,J Clin Microbiol.39:4456-4461;Houng,et al 2000,J Virol.86:1-11)。实时RT-PCR具有不同于噬斑检测和RT-PCR的优点,即它提供了快速的、定量的和特异的结果。
为尼帕病毒开发的TaqManTM检测法检测了大范围的病毒浓度,浓度为从每个反应1.2×105pfu到1.2pfu,它对应于200pfu/ml的阈值。对不同病毒的其他研究已经显示了相似的检测阈值(Houng,et al 2000,JVirol.86:1-11;Lanciotti,et al 2000,J Clin Microbiol.38:4066-4071)。发现尼帕病毒TaqManTM检测法的敏感度与那些用RT-PCR获得的敏感性相似(表2)。
TaqManTM检测法的可重复性是高的,因为在来自在不同时间以及用不同RNA制剂进行的一些检测的结果中,仅仅观察到了小的变异(见图7和表2)。因此,测试的可靠性可能主要依赖于RNA提取物。当使用尼帕病毒特异性引物和探针时,没有发生麻疹病毒RNA的扩增验证了尼帕病毒TaqManTM的特异性。麻疹病毒(Measles virus)是一种麻疹病毒(morbilivirus),它是与汉尼巴病毒最为紧密相关的属。最近已经开发出了亨德拉病毒的TaqManTM检测法,亨德拉病毒是一种在N基因上表现出与尼帕病毒有着78.4%核苷酸同源性的汉尼巴病毒(Smith,et al 2001,JVirol Methods.98:33-40;Wang,et al 2001,Microbes and Infection 3,279-287)。用程序Primer Express对尼帕病毒探针亲和力、亨德拉病毒N基因的前向和逆向引物的分析说明检测应当特异于尼帕病毒(Harcourt,et al2000,Virology.271:334-349)。用亨德拉病毒验证了尼帕病毒TaqMan检测法在汉尼巴病毒属中的特异性。用尼帕病毒特异性引物和探针时缺乏亨德拉病毒RNA的扩增确认了该测试对尼帕病毒的特异性。
RNA转录产物被开发作为定量检测法的稳定的、可重复的和可靠的标准物。尼帕病毒RNA定量的线性范围至少是109到103。得到了亨德拉病毒的相似结果:从未稀释的亨德拉病毒RNA到1/107中都观察到了线性(Smith,et al 2001,J Virol Methods.98:33-40)。这个线性范围容许检测大范围的病毒滴度,并应当可以定量地鉴定临床样品和细胞培养物中的尼帕病毒,而不需要稀释样品。令人奇怪的是,当在试管内稀释病毒时,增加了RNA分子/pfu的比值(表2),说明大量的RNA分子可能影响DNA扩增的效率。缺乏在含有大量RNA模板的样品中所具有的试剂也可能解释这一点。
发现用TaqManTM检测法计算出的病毒基因组分子的数目比用噬斑滴定测定出的感染病毒颗粒的相应数目高出约3个logs,对于登革热病毒,也发现每个感染pfu含有至少100或更多个基因组等价物,以及对于立夫特山谷热或普马拉病毒,注意到有2-3个log差异(Houng,et al2000,J Virol.86:1-11;Garcia,et al 2001,J Clin Microbiol.39:4456-4461;Garin,2001,Microbes Infect.3:739-745)。这个比例是因为非感染病毒的存在,或是因为缺陷的、未成熟的或灭活的颗粒的存在,或是因为如从受损的感染细胞中释放出的核颗粒的脱衣壳的RNA的存在。事实上,在感染后不同时间计算出的RNA/pfu比值随着感染时间而增加,在第4天观察到了最高比值,反映了细胞病变效应(图8)。
这些数据显示尼帕病毒TaqManTM检测法对于监测感染仓鼠的血清样品中的尼帕病毒也是有效的。在感染后第5天取得血清,因为这是唯一一天曾在动物中检测到病毒的时间(V.Guillaume et al.,J.Virol.2004.78:834-840)。但是,实时PCR和噬斑滴定都不能在5只仓鼠的两只仓鼠中证实有尼帕病毒,证实这些动物可能具有短暂的或不可检测的病毒血症。在仓鼠H3中检测到了病毒RNA而不是病毒。但是,仓鼠血清中的病毒滴度是相当低的,并接近于两种技术的检测低限(实时RT-PCR和噬斑滴度分别是200pfu/ml和100pfu/ml)。
实施例3:抗汉尼巴病毒的免疫接种和被动保护
在下面,已经在痘苗病毒重组体中表达出两种NiV糖蛋白(G和F)以评价它们对保护的贡献。为了实现这一点,使用了其中动物死于尼帕病毒感染后的急性脑炎的仓鼠动物模型,如实施例1所提出的(Wong etal.Am.J.Patol.2003.163:2127-2137)。利用这个模型,用表达两种尼帕病毒糖蛋白的其中一种的痘苗重组体免疫接种保护动物避免了致命性感染。此外,向纯真
动物被动转运免疫动物的抗体保护前者动物避免了致死性尼帕病毒攻击。
细胞和病毒
将Vero E6、RK13和BHK21细胞保持在含有10%胎牛血清的DMEM培养基(Gibco)中。在Vero细胞的两次传代之后,法国里昂的Jean Merieux BSL-4实验室收到了KB Chua博士和SK Lam博士(University of Malaya,Kuala Lumpur,Malaysia)的从患者的脑脊液中分离出的尼帕病毒。在Vero细胞的第3次传代之后,制备病毒原液(在P4条件下):在感染后第2天,当Veros细胞表现出融合及合胞体形成时,收集上清液。通过将每个孔内的200μl上清液的系列的10倍稀释液(每孔含有106个Vero细胞)在37℃下孵育1个小时,在6孔板中滴定病毒原液。然后用DMEM洗涤每孔细胞两次,并往每孔内加入2ml含有2%胎牛血清的1.6%羧甲基纤维素DMEM溶液。将板在37℃下孵育5天,用磷酸缓冲液(PBS)(pH 7.4)洗涤孔,并用10%福尔马林固定20分钟,洗涤并用亚甲兰染色。在以感染复数(MOI)为0.01感染Vero细胞之后,病毒滴度达到2×107pfu/ml。
将痘病毒和重组病毒的原液生长在BHK21细胞中。以0.01pfu/细胞感染细胞并在3天后收集、超声裂解细胞,储存在-80℃。在Vero细胞中滴定病毒。
NiV糖蛋白基因的克隆和痘苗重组体的构建
为了克隆编码两种病毒糖蛋白的NiV基因,根据产品说明,用RNA Now提取经NiV感染的Vero E6细胞,并进行RT-PCR。G蛋白所用的5’和3’引物分别是5’-CGCGGATCCAGTCATAACAATTCAAG-3’(SEQ ID NO:4)和5’-CGCGGATCCGAGGTTGATTTTTATG-3’(SEQ IDNO:5)。F蛋白的分别是5’-CGCAGGATCGAAGCTCTTGCCTCG-3’(SEQ ID NO:6)和5’-CATCAATCTGGATCCACTATGTCCC-3’(SEQ IDNO:7)。根据产品说明,将所形成的cDNA克隆到Pt-Adv质粒内。核酸序列分析发现,与已发表的NiV的核酸序列分析(Chan,et al 2001.J GenVirol.82:2151-5)比较,在NiV G基因的683位点有着单个核苷酸的差异(A到G),但是这种变化是沉默变化,基本序列是相关的。利用Perkus等(Perkus,et al 1989.J.Virol.63:3829-3836)描述的宿主范围选择系统制备VV重组体。简单地,通过用Bam HI从Pt-Adv质粒中切下插入物,并将其克隆到pCOPAK H6质粒的Bam HI位点而进行表达基因的亚克隆(Perkus,et al 1989.J.Virol.63:3829-3836),它也含有KIL痘苗基因。用VV的NYVAC株(Tartaglia et al 1992.Virology.188:217-232)感染以及用pCOPAK质粒转染Vero细胞。用RK13细胞选择VV重组体。
抗体确定
用酶联免疫吸附检测法(ELISA)个别地测定在感染仓鼠的血清中是否存在NiV抗体。从用MOI为0.01pfu/细胞感染24个小时的Vero细胞中制备出NiV抗原的粗提取物。用PBS洗涤细胞,并在4℃将其在含有1%Triton X100的PBS中裂解10分钟(107细胞/ml)。将细胞裂解液超声处理2次,每次30秒,以充分地破碎细胞,并将其在4℃、5000rpm离心10分钟。将上清液冻存在-80℃。相似地处理未感染的Vero细胞以制备抗原对照。用来自恢复期的、NiV感染患者的血清进行对尼帕病毒抗原的交叉滴定,以确定对应于表现为最高O.D.读数的稀释液的抗原滴度。
用Vero细胞确定中和抗体滴度。将血清PBS稀释液(开始于1/20)与50pfu NiV混和于96孔板中,并在37℃孵育1个小时,然后加入20,000个Vero细胞。在5天后读板,并记录下减少50%病毒滴度的血清稀释液。
引物和TaqManTM探针
所用的条件是如在实施例2上面所述的那些条件。简单地,按照所推荐的标准,用程序Primer ExpressTM(Perkin-Elmer,Applied Biosystems,USA)设计出引物和探针。选择NP基因的靶向区域。用前向引物(Ni-NP1209 5’GCAAGAGAGTAATGTTCAGGCTAGAG 3’-SEQ ID NO:1)和逆向引物(Ni-NP1314 5’CTGTTCTATAGGTTCTTCCCCTTCAT 3’-SEQID NO:2)扩增出了一个105个碱基对的NiV.NP基因。设计出荧光探针(Ni-NP1248Fam 5’TGCAGGAGGTGTGCTCATTGGAGG 3’-SEQ IDNO:3),其与在PCR引物内侧的序列退火。荧光报告染料6-羧基-荧光(FAM)位于探针的5’末端,以及猝灭剂6-羧基-四甲基-罗丹明(TAMRA)位于3’末端。
利用ABI PRISM 7700 TaqManTM系列检测仪进行定量RT-PCR检测。用一步RT-PCR系统(TaqManTM一步PCR基本混和试剂试剂盒,Applied Biosystems)进行一个连续的热循环。制备基本(master)混和反应物并将20μl部分或22.5μl部分分装到薄壁的microAmp光度管(ABIPRISMTM,Applied Biosystems)中。往每个管内加入5μl仓鼠血清的RNA提取物、或2.5μl原液病毒或感染细胞上清液的RNA提取物、或2.5μlRNA转录产物。最终反应混和物含有900nM每种引物和200nM每种探针。在扩增之前,将RNA在50℃下逆转录30分钟。接着是一个循环的94℃下变性5分钟,接下来,进行45个周期的94℃下15秒和60℃下1分钟的PCR扩增。
仓鼠的免疫接种
对于保护性研究,用107pfu表达G或F NiV糖蛋白的VV重组体或5×106个每种重组体(当它们用于共同免疫接种时)接种近亲杂交的金仓鼠(Janvier,Le Fenest St.Isles,France)两次(间隔1个月)。在末次接种后3个月,攻击动物。
为了制备多克隆的单特异性抗F和G糖蛋白血清,在第0天和第14天用107pfu VV重组体免疫接种仓鼠,随后在第28天用经超声处理的VV-重组体感染的BHK21细胞(+弗氏完全佐剂)以及在第42天用相同的抗原(+弗氏完全佐剂)免疫接种仓鼠。在末次免疫接种后第14天给动物取血,并用ELISA和中和法确定抗体。
NiV糖蛋白在痘苗病毒中的表达
对于生物活性蛋白的表达,在体外测试了NiV G或F蛋白在痘苗病毒中的表达。用FACScan分析检查了用VV-NiV.G或-F感染的HeLa细胞的NiV蛋白在细胞膜上的表达。两种病毒糖蛋白都表达于细胞膜上(图10)。当用两种痘苗重组体感染HeLa细胞时,诱导了细胞融合(合胞体形成)(图11)。
用表达G或F的VV重组体对仓鼠的免疫接种保护性对抗了致死性感染
用107pfu VV-NiV.G或F或两者的组合(5×106pfu每种重组体)皮下免疫接种仓鼠。1个月后,用相同剂量的痘苗重组体强化接种动物。在我们已经为NiV开发的动物模型中,用我们的NiV分离株对仓鼠的腹腔内接种7到10天之后诱导了致命性脑炎(见实施例1和图1)。当在末次免疫接种3个月后,用NiV攻击经VV-NiV.G、-F或G+F接种的动物时,有着对死亡率的完全保护(图12)。在攻击之后,在所有接种动物中都增加了如中和法及ELISA所检测的抗体的水平(图13)。对来自仓鼠的血清的其他研究显示在对照的非免疫接种的动物中,只有在感染的晚期(第5-6天)才能检测到病毒的存在。在接种动物中没有检测到病毒(表4)。
表4.定量分析对照血清和感染仓鼠血清中的NiV
以TaqMan assay*检测到的具有尼巴病毒RNA的仓鼠数量 | ||||
J1J2J3J4J5J6J7J8 | VV-NipG | VV-NipF | VV-NipG/VV-NipF | 对照 |
-(4)--(4)--(4)--(4)- | -------- | -------- | ----4(5)2(3) |
*对于每一个免疫接种测试,每天测试5只动物
来自VV-NiV.G或-F重组体免疫接种的仓鼠的血清被动地保护了纯真仓鼠对抗致死性NiV攻击。
为了细剖体液免疫应答在保护中的重要性,用痘苗重组体超免疫仓鼠(见材料和方法),并汇聚具有含高水平的NiV的中和抗体的血清的动物(160中和单位/ml)。经腹腔内注射给予仓鼠0.2ml针对性对抗G或FNiV糖蛋白或两者的混和物的抗血清。1个小时之后,用病毒攻击动物,并在24个小时之后再次被动转运0.2ml血清。在两个月内观察仓鼠的临床征象。接受抗血清(单特异的多克隆G或F)或两者的混和物的动物能避免致死性NiV感染(图14)。在感染后,强烈地诱导了抗NiV的血清抗体水平(图15)。
上述显示了可能在保护性对抗NiV感染中起到作用的免疫学参数。
用VV.G或G接种的仓鼠能完全避免致死性感染。确认了体液应答在这个过程中的贡献,也显示通过在攻击之前被动转运超免疫血清也能保护纯真动物。因此,利用动物模型,上述显示主动地或被动地保护性对抗致死性NiV感染是可能的。但是,在主动和被动免疫中都强烈地刺激了针对NiV的抗体应答,说明病毒在接种动物体内复制。但是,尝试检测血清中的病毒还不成功。在对照的非免疫接种的动物中,仅能在濒死动物的血清中检测到病毒。如在一些其他的副粘病毒感染中所观察到的那样,可能是因为病毒主要是细胞相关性病毒。
在人体中,已经观察到了感染的复发性和迟发性病例(Lim,et al2003.J.Neurol.Neurosurg.Psychiatry.74:131-133;Tan,et al 2002.AnnNeurol.51:703-708;Wong,et al 2001.J Neurol Neurosurg Psychiatry.71:552-554)。在这些情况中,还不知道感染的免疫生物学。在我们的受攻击的免疫接种的动物中,直到攻击后5个月还没有观察到这些迟发性病理。相似地,在被动保护的动物中,也没有观察到迟发性疾病。但是,还没有确定出在体内的抗体保护的低限或者一旦已经开始被感染时被动免疫动物的作用。
显而易见的是,根据上述教导,对本发明作出各种修饰和变异都是可能的。因此,要明白只要在所附的权利要求书的范围之内,可以以不同于在此所具体描述的方式实践本发明。
实施例4:抗尼帕病毒的单克隆抗体的生产和反应性
为了研究尼帕病毒感染的病理,我们已经建立了仓鼠模型(权利要求书部分)。在用尼帕病毒感染之后,动物死于脑炎,其表现出与在人体中所见相似的病理。此外,我们已经显示通过利用糖蛋白(G或F)的疫苗接种或利用针对性对抗其中一种抗原的抗血清被动地保护了这些动物(权利要求书部分)。虽然至今还没有得到一种汉尼巴病毒感染的治疗方法,但是我们将开发出一种免疫治疗方法,为汉尼巴病毒感染个体发生预防作用。
我们已经开发了一个针对NiV G和F糖蛋白的单克隆抗体(mAb)库,它在体外中和尼帕病毒的传染性。此外,某些抗NiVF mAb可中和亨德拉病毒。
现状和可用材料
我们已经从所制备的抗表达G或F的尼帕病毒蛋白的抗体库中特征性地描述出了30种mAb。17种抗NiF以及13种抗NiG。根据病毒中和作用,已经选择出一些用于本研究。应当注意,没有一种抗NiG中和亨德拉病毒,而抗NiF也中和HeV。已经用竞争性ELISA以及也通过测序逃逸突变体(escape mutants)研究了这些NiV mAb所识别的抗原表位。下面显示了选择用于最初研究的mAb的性能:
抗原 | 名称 | 同种型 | 识别氨基酸(逃逸突变体) | 中和特异性 | |
NiV | HeV | ||||
GGGGFF | 1.73B105A77F3353 | IgG1IgG1IgG2aIgG2bIgG1IgG2a | 336,391500,533n.d.n.d.282(NiV),216(HeV)247(NiV&HeV) | 1.7×1060.5×1061.1×1051.3×1053.5×1052.4×105 | ----3.5×1051.2×105 |
对于NiV感染后的免疫应答的分析,我们已经在痘苗病毒中表达了G、F和NP NiV蛋白。在ELISA检测中,用这些从感染细胞的裂解物中得到的抗原测定抗原特异的应答。
用含有尼帕病毒G或F蛋白的cDNA的表达质粒VIJ免疫接种Balb/c小鼠。利用基因枪(BioRad)技术已经实现了这一点。已经用编码尼帕病毒G或F蛋白的痘苗重组体强化接种小鼠,并在此次强化接种后3-4个月,在融合前3天,给小鼠照射经照射的尼帕病毒感染的Vero细胞。用尼帕病毒感染的和未感染的Vero细胞筛选分泌IgG杂交瘤的杂交瘤。
我们通过中和作用已经特征性地描述了所有的NiV mAb,以及通过竞争性ELISA和对逃逸突变体的序列分析描述了mAb的数目。总而言之,我们的研究至今发现在F或在G蛋白上可能有着一个单一的主要的识别表位,以及逃逸突变体的数据说明不同的mAb不同程度地重叠着该区域。
序列表
<110>巴斯德研究院
国家健康与医学研究院
<120>尼帕病毒检测方法和提供抗汉尼巴病毒的免疫保护的方法
<150>US 60/584,472
<151>2004-07-02
<150>US 60/504,225
<151>2003-09-22
<160>45
<170>PatentIn version 3.2
<210>1
<211>26
<212>DNA
<213>Artificial Sequence
<220>
<223>synthetic oligonucleotide
<400>1
gcaagagagt aatgttcagg ctagag 26
<210>2
<211>26
<212>DNA
<213>Artificial Sequence
<220>
<223>synthetic oligonucleotide
<400>2
ctgttctata ggttcttccc cttcat 26
<210>3
<211>24
<212>DNA
<213>Artificial Sequence
<220>
<223>synthetic oligonucleotide
<400>3
tgcaggaggt gtgctcattg gagg 24
<210>4
<211>26
<212>DNA
<213>Artificial Sequence
<220>
<223>synthetic oligonucleotide
<400>4
cgcggatcca gtcataacaa ttcaag 26
<210>5
<211>25
<212>DNA
<213>Artificial Sequence
<220>
<223>synthetic oligonucleotide
<400>5
cgcggatccg aggttgattt ttatg 25
<210>6
<211>24
<212>DNA
<213>Artificial Sequence
<220>
<223>synthetic oligonucleotide
<400>6
cgcaggatcg aagctcttgc ctcg 24
<210>7
<211>25
<212>DNA
<213>Artificial Sequence
<220>
<223>synthetic oligonucleotide
<400>7
catcaatctg gatccactat gtccc 25
<210>8
<211>18246
<212>DNA
<213>Nipah virus
<400>8
accaaacaag ggagaatatg gatacgttaa aatatataac gtatttttaa aacttaggaa 60
ccaagacaaa cacttttggt cttggtattg gatcctcaag aaatatatca tcatgagtga 120
tatctttgaa gaggcggcta gttttaggag ttatcaatct aagttaggga gagatgggag 180
ggctagtgca gcaactgcta ctttgacaac caagataagg atatttgtac cagctactaa 240
tagtccagag ctcagatggg aactaacatt gtttgcactt gatgtgatta gatctccgag 300
tgctgccgag tcaatgaaag ttggagctgc tttcacactc atctctatgt attcagagag 360
acccggggct ctcattagaa gtctcctcaa tgacccagac attgaagctg taataataga 420
tgttggatca atggtcaacg gaataccagt aatggagagg agaggagaca aggctcagga 480
ggagatggaa ggcttgatga gaatcctcaa aactgctcga gacagcagca agggaaaaac 540
accttttgtt gacagccgag cttacggcct acggataaca gacatgagca ccctggtctc 600
tgcagttatc accatcgagg cccagatctg gatactgatc gctaaagcag ttacagctcc 660
cgacactgcc gaggaaagtg aaactagaag atgggctaaa tacgtccaac aaaagagagt 720
caatccgttc tttgctctaa ctcagcaatg gctaacagaa atgaggaatc tgctctccca 780
gagtctatca gtaaggaagt tcatggttga gatcctcata gaagtcaaga aaggaggatc 840
tgctaaaggc agagcagtag aaataatctc agacatcgga aactatgtcg aggaaactgg 900
tatggcagga ttcttcgcaa ccatcagatt cgggttggag acaaggtatc cagcacttgc 960
actcaacgaa ttccagagtg acctcaacac catcaaaagc ttgatgctac tctacagaga 1020
aattggccca agagcccctt atatggtgct tcttgaagaa tcaattcaga ctaaatttgc 1080
ccctggaggt tacccattat tgtggagctt tgccatgggt gtggctacta ctattgacag 1140
gtctatgggg gcattgaata tcaatcgtgg ttatcttgag cctatgtatt tcagactagg 1200
ccaaaaatca gcacgtcacc atgctggagg aattgatcag aacatggcaa atagactggg 1260
actaagttca gatcaagttg cagaactcgc tgctgcagtt caggaaacat cagcaggaag 1320
gcaagagagt aatgttcagg ctagagaggc aaaatttgct gcaggaggtg tgctcattgg 1380
aggcagtgat caagatatcg atgaagggga agaacctata gaacagagtg gcagacagtc 1440
agttaccttc aaaagggaga tgagtatttc atcccttgct aacagtgtgc cgagcagttc 1500
tgtgagcaca tccggtggga ccagattgac taattcatta ctaaacctca gatcaagact 1560
ggctgcaaaa gcagcaaaag aagccgcctc atccaatgca acagatgatc cagcaatcag 1620
caacagaact caaggggaat cagagaagaa gaataatcaa gacctcaaac ctgctcaaaa 1680
tgaccttgat ttcgtcagag ctgatgtgtg acgtctattt ccaatattct acagtatcca 1740
aaaatctttc tatagtacac tatcataata cgacactaag ggatcaacca tatcaaagtt 1800
acgaatcgtt ttaattatat taatcaaatg atactctttt atgggcaaac cgaagaacca 1860
atgtctacat gtaaattgag ctttggtatt gcaatctaat acttgctcaa aatcttgaac 1920
tattagtgta atttctatca tcatagagtt atcaagattt tattatataa gttggtgcag 1980
atctttggac atgaattaca cactacactc taatgaagac aaaatttaca ttacatattt 2040
aaggactatt tcctatcctt tcaatggtac ttggttatga aggtttctta atttaactaa 2100
gctactgtct ttgcactgga atatacaata cctcttacct catttcttac tttaatatca 2160
tgttattttt ttgataagtc acttaacttg accaaggtct accaggtaat gctcgcacaa 2220
gtgaactgca atctcaactt agattaaaca taatcatgca aaatcactat tttgtactac 2280
taactcatta agaaaaactt aggatccaag agatttactc taggatctcc tattaagctt 2340
agcagtcatt agttgagagt tcaacttgca aaactctaac cttcactcta ataacaattc 2400
atccaatgga taaattggaa ctagtcaatg atggcctcaa tattattgac tttattcaga 2460
agaaccaaaa agaaatacag aagacatacg gacgatcaag tattcaacaa cccagcatca 2520
aagatcaaac aaaagcctgg gaagattttc tgcagtgcac cagtggagaa tctgaacaag 2580
ttgagggggg aatgtctaag gatgatggag atgttgaaag aagaaacttg gaggatctat 2640
ccagtacttc tcccacagat ggaactattg gaaagagagt gtcgaacacc cgtgactggg 2700
cagaaggttc agatgacata caactggacc cagtggttac agacgttgta taccatgatc 2760
atggaggaga atgtaccgga tatggattta cttcaagccc tgagagaggg tggagtgatt 2820
acacatcagg agcaaacaat gggaatgtat gtcttgtatc tgatgcaaag atgctgtcct 2880
atgctcccga aattgcagtt tctaaagaag atcgggaaac tgatctagtt catcttgaga 2940
ataaactatc tactacagga ctgaatccca cagcagtacc gttcactctg agaaacctgt 3000
ctgatcctgc aaaagactct cctgtgattg ctgaacacta ctacggacta ggagttaaag 3060
agcaaaacgt tggccctcag actagcagaa atgtcaattt ggacagcatc aaattgtaca 3120
catcagatga cgaagaggca gatcagcttg aattcgaaga tgagtttgca ggaagctcaa 3180
gtgaagtgat agtcggcatt tctcctgaag atgaagagcc ttcaagtgtt ggcggaaaac 3240
ccaatgaatc cattggacgt acaatcgaag gccaatcaat ccgagacaac cttcaagcca 3300
aggacaacaa atcaacagat gtaccaggag caggaccgaa agattcagca gtgaaggaag 3360
aaccacccca gaagaggcta cctatgttag ctgaagaatt tgagtgctct ggatcggaag 3420
acccaatcat tcgggagctg ctgaaggaga actcactcat aaattgtcag caagggaaag 3480
atgctcagcc tccatatcat tggagcatcg agaggtcaat aagcccggat aaaactgaga 3540
tcgtcaacgg tgctgtgcaa actgctgaca ggcaaagacc aggaactccg atgccaaagt 3600
cccgaggtat tcccattaaa aagggcacag acgcgaaata tccatctgct gggacggaaa 3660
acgtgcctgg gtcgaagagt ggtgcaaccc ggcatgttcg aggatcaccc ccctaccaag 3720
aaggcaagag tgtcaatgcg gagaatgtcc aactgaatgc ttccactgcg gttaaggaaa 3780
ctgataagtc agaagtaaac cccgtagacg acaacgactc acttgatgat aaatacatca 3840
tgccttcaga tgatttctca aacactttct tcccgcacga cactgatcgc ttgaattatc 3900
acgcagatca tttaggtgat tatgaccttg aaaccctgtg tgaagagtcg gttctaatgg 3960
gagtgatcaa ctctataaaa ttaattaatc tggatatgcg cttaaatcac attgaagaac 4020
aagttaaaga gatcccaaag atcatcaata agcttgagtc cattgacaga gttctggcca 4080
agactaacac cgcactctca accattgaag gacacctggt ttccatgatg ataatgatac 4140
cagggaaagg gaaaggagaa agaaagggga aaaataatcc tgagcttaaa ccagtgatag 4200
gaagagacat tctagagcag caatctcttt tttcttttga caatgtcaag aatttcagag 4260
atggatcgtt gacaaacgaa ccgtatgggg cagctgtaca gttgagagaa gatcttattc 4320
ttcctgaact taattttgag gagacaaatg catctcaatt tgttcctatg gcagatgatt 4380
catccagaga tgttatcaag acattgataa ggactcacat taaagataga gagttgagat 4440
cagaactgat tggttacctg aataaagcgg aaaatgatga ggaaattcag gagatagcga 4500
acactgtcaa tgacatcatt gacggtaata tttgatcact gaattgtcag cagaaataca 4560
atgatctaac aacaatctcc cacaagtaga caatggtttc aggtcaataa taacaacctc 4620
aatactaatc tttcacataa gcattactca ttccagccct cagacgataa cacaatactt 4680
gatacatgtt tattgaagtg tatgtagcat gattgaacta ttcaataact gtatttctca 4740
ctcttgctct tagttagtca ttgtgtctaa taattattat tacagtacaa ggtattatga 4800
attcaaagat acgcaataaa tctgatatca gcatagagta gaaaattgtt gtttttgtca 4860
tgatcattcg aagatttaac aatgatgtca actttcatac ctaaacataa taacataaaa 4920
tggtcgattt gtattgtaga tctctcacgc attttagtgt catgaattag tgtttcaaat 4980
cagttgcata tcaattaaga aaaacttagg agacaggtat agaacctctc tttcagataa 5040
ctggtcaatt aaggacagaa attctgtttc tcaaatccgc tagcctttgt caaagaggac 5100
acaagcaatg gagccggaca tcaagagtat ttcaagtgag tcaatggaag gagtatctga 5160
tttcagccct agttcttggg agcatggtgg gtatcttgat aaggttgaac cagaaattga 5220
tgaaaatggc agtatgattc caaaatacaa gatctatacc ccaggagcta acgagaggaa 5280
atacaacaac tacatgtacc ttatatgtta cggctttgtt gaagatgttg agagaacccc 5340
agagacaggg aaacgcaaga agatcaggac aattgctgcc taccctctgg gtgttggtaa 5400
gagtgcctct catccccaag atcttctgga ggaactctgt tccctcaaag ttactgtgag 5460
aagaacagct ggatcaactg agaaaattgt gtttggatca tctggccctc taaatcacct 5520
cgttccgtgg aagaaagtac tgactagtgg ttcaattttt aatgcagtca aggtttgtcg 5580
gaacgttgat cagatacagc ttgacaagca tcaagctctg agaatatttt ttctcagtat 5640
cacaaagctc aatgattctg gaatctacat gattccacga accatgcttg agttcaggag 5700
aaacaatgcc attgccttca atcttctagt gtacttgaag attgatgctg atttatccaa 5760
aatggggatc cagggaagcc tcgataaaga tggcttcaag gttgcctcct tcatgctaca 5820
cttggggaac tttgtccgtc gtgcagggaa gtattactct gttgattatt gtaggaggaa 5880
gattgatagg atgaaattgc agttttcact gggttccata ggcggactaa gtctccacat 5940
taagatcaat ggtgtaatca gcaaacggct gtttgctcaa atgggattcc aaaaaaacct 6000
ttgtttctct ttgatggaca tcaatccttg gctcaacaga ttgacctgga acaacagttg 6060
tgagatcagc cgagtagcag ctgtgttgca gccttctatt ccaagagagt tcatgatcta 6120
tgatgatgtc ttcattgaca atacagggag aattctaaag ggctaaacag aattcttcta 6180
aaatttaatc agtcatgagt ttagtaatca tacctagtca taatacatca cacaggacta 6240
tttacaaaag acagttaaaa aatggaataa tcatgtagta gtaattgaga acattattag 6300
aatagtataa ctaaaatgta gtttttttga gtatttgatt taaaattaga taactattac 6360
aaaaaactta ggagccaagc tcttgcctcg ttcagaaggt taaacaagca ttcttaccat 6420
tggatcaaca aaaggattgg ttttatcgtc taagaaattt attgaaaggc aaagaaattc 6480
ctggttttat gttgaatgag gtgtatcaaa ctaaggagac cttctaacag ccaggtcata 6540
ggaatataaa taaaaataag aataaaattg attccatcgg aagattcatt tcaagaagtg 6600
atcaaatcaa agcggttggc agacctacca atcatatacc acaagactcg acaatggtag 6660
ttatacttga caagagatgt tattgtaatc ttttaatatt gattttgatg atctcggagt 6720
gtagtgttgg gattctacat tatgagaaat tgagtaaaat tggacttgtc aaaggagtaa 6780
caagaaaata caagattaaa agcaatcctc tcacaaaaga cattgttata aaaatgattc 6840
cgaatgtgtc gaacatgtct cagtgcacag ggagtgtcat ggaaaattat aaaacacgat 6900
taaacggtat cttaacacct ataaagggag cgttagagat ctacaaaaac aacactcatg 6960
accttgtcgg tgatgtgaga ttagccggag ttataatggc aggagttgct attgggattg 7020
caaccgcagc tcaaatcact gcaggtgtag cactatatga ggcaatgaag aatgctgaca 7080
acatcaacaa actcaaaagc agcattgaat caactaatga agctgtcgtt aaacttcaag 7140
agactgcaga aaagacagtc tatgtgctga ctgctctaca ggattacatt aatactaatt 7200
tagtaccgac aattgacaag ataagctgca aacagacaga actctcacta gatctggcat 7260
tatcaaagta cctctctgat ttgctttttg tatttggccc caaccttcaa gacccagttt 7320
ctaattcaat gactatacag gctatatctc aggcattcgg tggaaattat gaaacactgc 7380
taagaacatt gggttacgct acagaagact ttgatgatct tctagaaagt gacagcataa 7440
caggtcaaat catctatgtt gatctaagta gctactatat aattgtcagg gtttattttc 7500
ctattctgac tgaaattcaa caggcctata tccaagagtt gttaccagtg agcttcaaca 7560
atgataattc agaatggatc agtattgtcc caaatttcat attggtaagg aatacattaa 7620
tatcaaatat agagattgga ttttgcctaa ttacaaagag gagcgtgatc tgcaaccaag 7680
attatgccac acctatgacc aacaacatga gagaatgttt aacgggatcg actgagaagt 7740
gtcctcgaga gctggttgtt tcatcacatg ttcccagatt tgcactatct aacggggttc 7800
tgtttgccaa ttgcataagt gttacatgtc agtgtcaaac aacaggcagg gcaatctcac 7860
aatcaggaga acaaactctg ctgatgattg acaacaccac ctgtcctaca gccgtactcg 7920
gtaatgtgat tatcagctta gggaaatatc tggggtcagt aaattataat tctgaaggca 7980
ttgctatcgg tcctccagtc tttacagata aagttgatat atcaagtcag atatccagca 8040
tgaatcagtc cttacaacag tctaaggact atatcaaaga ggctcaacga ctccttgata 8100
ctgttaatcc atcattaata agcatgttgt ctatgatcat actgtatgta ttatcgatcg 8160
catcgttgtg tatagggttg attacattta tcagttttat cattgttgag aaaaagagaa 8220
acacctacag cagattagag gataggagag tcagacctac aagcagtggg gatctctact 8280
acattgggac atagtgtatt cagattgatg aaattatgtt agagaaatca gaaaacttct 8340
gactttcaga aatggattgt atacaattag ttagatcatc ctgaataatc gaggtgagaa 8400
cattgcaact ataaaatcag atcatgtaaa tagttgtaaa aaattaaaag cttcttttaa 8460
ttcttttgaa caataattta attaatatat aacatattct ctcacacgag cgctaaccta 8520
tacactctct actaatattt tatactcata attaatgata taatgacaaa taaggattca 8580
aattggatta tgatatagtt tcatactaca atagcatttc gaccaagaaa atatccttac 8640
aattatacaa tgtacttaac cgtgaatatg taattgataa tttcccttta gaaatttaat 8700
aaaaaactta ggacccaggt ccataactca ttggatactt aactgtatct ttctaagcta 8760
tcacatatca aaggagagat tgaatgcttt tttggagatc tagatcatta ctatatgtgt 8820
ctcctataat cacatcatag gagtgaacca taatacacat ctttgggtag gggaaggaaa 8880
gtattgttga cgtactgatt gatctgcttg agtcaaataa tcagtcataa caattcaaga 8940
aaatgccggc agaaaacaag aaagttagat tcgaaaatac tacttcagac aaagggaaaa 9000
ttcctagtaa agttattaag agctactacg gaaccatgga cattaagaaa ataaatgaag 9060
gattattgga cagcaaaata ttaagtgctt tcaacacagt aatagcattg cttggatcta 9120
tcgtgatcat agtgatgaat ataatgatca tccaaaatta cacaagatca acagacaatc 9180
aggccgtgat caaagatgcg ttgcagggta tccaacagca gatcaaaggg cttgctgaca 9240
aaatcggcac agagataggg cccaaagtat cactgattga cacatccagt accattacta 9300
tcccagctaa cattgggctg ttaggttcaa agatcagcca gtcgactgca agtataaatg 9360
agaatgtgaa tgaaaaatgc aaattcacac tgcctccctt gaaaatccac gaatgtaaca 9420
tttcttgtcc taacccactc ccttttagag agtataggcc acagacagaa ggggtgagca 9480
atctagtagg attacctaat aatatttgcc tgcaaaagac atctaatcag atattgaagc 9540
caaagctgat ttcatacact ttacccgtag tcggtcaaag tggtacctgt atcacagacc 9600
cattgctggc tatggacgag ggctattttg catatagcca cctggaaaga atcggatcat 9660
gttcaagagg ggtctccaaa caaagaataa taggagttgg agaggtacta gacagaggtg 9720
atgaagttcc ttctttattt atgaccaatg tctggacccc accaaatcca aacaccgttt 9780
accactgtag tgctgtatac aacaatgaat tctattatgt actttgtgca gtgtcaactg 9840
ttggagaccc tattctgaat agcacctact ggtccggatc tctaatgatg acccgtctag 9900
ctgtgaaacc caagagtaat ggtgggggtt acaatcaaca tcaacttgcc ctacgaagta 9960
tcgagaaagg gaggtatgat aaagttatgc cgtatggacc ttcaggcatc aaacagggtg 10020
acaccctgta ttttcctgct gtaggatttt tggtcaggac agagtttaaa tacaatgatt 10080
caaattgtcc catcacgaag tgtcaataca gtaaacctga aaattgcagg ctatctatgg 10140
ggattagacc aaacagccat tatatccttc gatctggact attaaaatac aatctatcag 10200
atggggagaa ccccaaagtt gtattcattg aaatatctga tcaaagatta tctattggat 10260
ctcctagcaa aatctatgat tctttgggtc aacctgtttt ctaccaagcg tcattttcat 10320
gggatactat gattaaattt ggagatgttc taacagtcaa ccctctggtt gtcaattggc 10380
gtaataacac ggtaatatca agacccgggc aatcacaatg ccctagattc aatacatgtc 10440
cagagatctg ctgggaagga gtttataatg atgcattcct aattgacaga atcaattgga 10500
taagcgcggg tgtattcctt gacagcaatc agaccgcaga aaatcctgtt tttactgtat 10560
tcaaagataa tgaaatactt tatagggcac aactggcttc tgaggacacc aatgcacaaa 10620
aaacaataac taattgtttt ctcttgaaga ataagatttg gtgcatatca ttggttgaga 10680
tatatgacac aggagacaat gtcataagac ccaaactatt cgcggttaag ataccagagc 10740
aatgtacata aaaatcaacc tcataattta atggattgat ctaatataat gataataatc 10800
gtacaaagac atgtgatgta aacaaaattg ttgtaattaa ataagtcctc agctgaatac 10860
ttttttaaga ttagcaatag catgtttttc cagttattgg atagttgata atataattct 10920
gaaactgggt taataaataa tcttgatcgg tgatctttga gaacaatgat atcatatagt 10980
tcatcaagtg ataatcaatt ctttatatgt acactttaga gtatattttg agacttagta 11040
ttttcggccc gaatgttaaa tttaatagtt catacataac ctaaactcaa gttctaagca 11100
taatgataac aattaatgcg aacttgtctt gatgtaagga agatttgata ttaactgaga 11160
ctccacttga tatagtagag ctgaatcttg taaataaatt ataatgaata gtttattcaa 11220
agattatcat tcatattagt gtaaattaag aaaaacttag gacccaggtc cttgattatg 11280
ccaattttct cgagaaatca ttcaattgac catagactga aagcgttgtt acctagttct 11340
tcagaagaga tcttattaga attaatttat atgatctaat tcccttaaaa actgaatacc 11400
aaaaaacaaa aatggccgat gaattatcaa tatccgacat catttaccct gaatgtcatt 11460
tggatagtcc tatagtctct ggtaaactaa tatcagctat tgaatatgct caattgagac 11520
acaatcagcc cagtgatgat aaaagactgt ctgagaatat taggttaaac cttcacggga 11580
aaagaaagag tctatacata ttaagacaat ccaaacaggg tgattacatt agaaacaaca 11640
taaaaaacct aaaggaattc atgcatattg cgtaccctga atgcaataac attctattct 11700
ccatcacatc ccaaggcatg actagcaaac ttgataacat catgaaaaag tcattcaaag 11760
catacaatat cattagtaag aaagtaattg ggatgctgca aaatatcact agaaatctca 11820
taactcaaga tagaagagat gaaataatta atatacatga gtgtaggcga ttaggggatt 11880
tagggaagaa tatgagtcaa tctaaatggt atgagtgttt tttgttttgg tttactatca 11940
aaacagagat gcgagcagtg atcaagaatt cgcaaaagcc gaaattccgt tcagattcat 12000
gcataataca catgcgagac aaaagtactg aaataatcct aaatccgaat cttatctgca 12060
ttttcaaatc agacaaaact ggaaagaagt gttattatct tacacccgaa atggttctaa 12120
tgtattgtga tgtcctagag ggaaggatga tgatggagac aacagtcaaa tcggatatca 12180
agtaccaacc tctaatctcg agatccaatg ccctctgggg gctaattgat cccttgttcc 12240
ctgtcatggg aaacagaatt tacaatatag tgtctatgat agagccttta gttcttgcac 12300
tactccaact caaggatgag gctaggatcc tgaggggtgc atttctgcat cactgcataa 12360
aggaaatgca tcaagaattg agtgagtgtg gttttacaga tcagaagatt cggtctatgt 12420
ttattgatga tcttttatcc attctaaata tcgataatat acatctgttg gcagagttct 12480
tttctttctt tcgtacgttt ggccatccta ttcttgaggc taaagttgct gcagaaaaag 12540
tgagagaaca tatgttggca gataaagttc ttgaatatgc ccctataatg aaagcacatg 12600
ctatattctg cgggactata ataaatgggt atagggatag acacggagga gcctggcctc 12660
ctctttacct ccccgcacat gcatctaaac atataatccg tttgaaaaat tctggggaat 12720
ctttgaccat tgatgactgt gtcaagaatt gggaatcatt ctgtgggatt caatttgatt 12780
gtttcatgga gctgaaattg gacagtgatc tgagtatgta tatgaaagat aaagctttat 12840
ctccaatcaa agacgaatgg gacagtgtat acccacgtga agtgttgagc tataccccac 12900
cgaagtcaac cgagccaaga agattggttg acgtttttgt aaatgatgaa aactttgatc 12960
catacaacat gctggaatat gtcttatccg gtgcttatct cgaggatgaa caattcaatg 13020
tttcttatag cttgaaggag aaagagacga agcaagctgg acgattgttc gcaaagatga 13080
cctacaaaat gcgtgcatgt caagtcatag cagaggccct gatagcctca ggtgtcggta 13140
aatattttaa ggagaacggg atggttaagg atgagcacga acttttgaag acactcttcc 13200
aattgtctat ttcctcagtt cctcgaggga acagtcaggg taatgatcct caatccatca 13260
ataatataga aagagatttc caatacttta aaggggtcac taccaatgtg aaagacaaaa 13320
agaataactc ttttaataag gttaaatctg ctctcaataa tccgtgccaa gctgacggag 13380
tccatcataa catgtcaccc aatacacgaa atcgttataa gtgtagtaat acaagtaagt 13440
cttttctcga ttatcatacc gagtttaatc ctcacaatca ctataaatca gacaatacag 13500
aggcggccgt actgtccagg tatgaggaca acactgggac aaaatttgat acagtaagtg 13560
catttcttac aactgatctt aagaaattct gtctcaattg gagatacgaa tcaatggcta 13620
tatttgctga acgtctggat gagatatacg gtttacctgg attttttaat tggatgcaca 13680
aacgactaga aagatctgtt atctatgttg cagaccctaa ttgcccccct aatattgaca 13740
aacatatgga actagaaaaa actcctgaag atgatatatt cattcattat cctaaaggcg 13800
gtattgaagg atatagccaa aaaacatgga ctatagcaac tatccccttt ttattcttga 13860
gtgcctatga gacaaacacg aggattgctg caattgtcca aggagacaat gaatcaattg 13920
ctatcactca aaaagttcat cctaatcttc cctacaaggt aaagaaagag atctgtgcaa 13980
agcaagctca gctttatttt gaaaggttaa ggatgaactt aagagccctc ggccacaatc 14040
ttaaagctac agaaactatc atcagtacac atctttttat ttattcgaag aaaattcatt 14100
atgatggtgc tgtgctgtct caggcactca aatcaatgtc aagatgttgc ttttggtcag 14160
agactctggt ggatgaaact agatcagctt gtagtaacat cagcactaca atagctaaag 14220
ctatagaaaa tgggttgtca agaaatgtcg gctattgcat caatattttg aaagtaattc 14280
agcagcttct catatcaact gagtttagta ttaacgagac attgacactg gatgtgacat 14340
ctcccatttc aaataattta gattggctta taacagctgc attaatcccg gcacctattg 14400
gaggattcaa ttaccttaat ttgtctagaa tttttgttag aaatataggt gatccggtta 14460
cagcatcttt ggctgatctt aagagaatga ttgatcacag tattatgact gaaagcgtat 14520
tacaaaaagt tatgaatcaa gaacctggtg atgcgagttt cttggactgg gccagtgatc 14580
catactcggg caacttgcct gactcacaaa gcatcactaa aacaattaaa aatatcacag 14640
caaggactat actgaggaac tcaccgaacc caatgctaaa aggtttattt catgacaaat 14700
cttttgatga agatcttgaa ctagctagct tcttaatgga caggagggtt atattaccta 14760
gagccgctca tgagatactg gataattcat tgacaggtgc cagagaggaa attgctggtt 14820
tattagatac aactaaaggc ttgatcagat cagggctaag aaagagtgga cttcagccaa 14880
agttagtttc tagattatct catcatgatt ataatcaatt tttaatactg aacaaacttc 14940
tatcaaacag aagacaaaat gacttgatat catcaaatac ttgctcagtt gacttggcac 15000
gagcattgag atctcacatg tggagggaat tagcgttagg tagagtaata tacggtcttg 15060
aggtaccaga tgcacttgag gctatggtgg gaaggtatat aacagggagc ttagagtgcc 15120
aaatttgtga gcagggaaac acgatgtatg ggtggttctt tgtacctagg gattcccaat 15180
tggatcaggt agatagagag cactcatcaa taagagtacc ttatgtagga tcaagtacgg 15240
atgaaagatc ggatatcaaa ctagggaatg tcaaaagacc aactaaggcc ttgcgttctg 15300
ctatcagaat tgcgacagta tatacttggg cctatgggga caatgaagag tgttggtatg 15360
aagcttggta cctagcgtct cagagggtaa acatagactt agatgtattg aaagctataa 15420
ccccagtttc cacttcaaac aatttatccc atagattgag agataaatcc acacaattta 15480
agtttgcagg gagtgtactc aacagagttt ctagatatgt taacataagc aatgacaatc 15540
tagatttcag aattgaggga gaaaaggtag atacgaatct tatttatcaa caagcaatgc 15600
tattagggtt atcggtattg gaaggtaaat tcagattgag attagaaact gatgattaca 15660
acgggatata tcacttacac gtaaaggata attgttgtgt caaagaagtg gctgatgtag 15720
gccaagtaga cgctgagttg cctatcccag aatatactga agtggataac aatcatctta 15780
tatatgatcc agaccccgtt tcagaaatag attgcagccg tctttctaat caggagtcca 15840
aatcaagaga attagacttt cctttatggt caactgagga acttcatgat gtcctagcta 15900
agactgttgc tcagaccgtt cttgagatta taacaaaggc tgacaaggat gttttaaagc 15960
aacaccttgc aatagactct gacgataaca tcaacagctt aatcacagaa tttctaatag 16020
ttgatcctga actgtttgca ctttatctag gacaatctat atcaataaaa tgggcctttg 16080
aaattcatca taggcgtcct agaggaagac atactatggt cgacctattg tcagatcttg 16140
tatcaaatac atcaaagcac acttacaaag tgttgtcaaa tgccttgtca catcctagag 16200
tattcaagag atttgtaaac tgtggcttgc tattgcctac acagggtcct taccttcatc 16260
aacaagattt tgaaaagttg tctcaaaacc ttcttgtaac atcttatatg atttatctaa 16320
tgaactggtg tgacttcaag aaatccccct ttttaatcgc cgaacaggat gaaactgtga 16380
taagtctacg agaggatata ataacatcca aacatctctg tgttataatt gacttatatg 16440
caaatcacca taaacctcct tggataatag atctaaaccc acaagaaaaa atatgtgtac 16500
tgcgtgactt tatttctaaa tctaggcatg tggacacgtc ctccagatca tggaatactt 16560
ctgacctgga ttttgtaata ttctatgcat ctttgactta tttgagaaga ggtataataa 16620
aacaattaag gataagacaa gttactgagg ttatagatac cacaacaatg ttaagggaca 16680
atataattgt agagaatcct cctattaaaa caggagtgtt agacatcaga ggttgtataa 16740
tatacaattt agaggaaatc ctgtctatga acacaaaatc agcatcaaaa aagatcttta 16800
atcttaatag taggccgtca gtggagaatc ataaatatag aaggataggt ctcaactcat 16860
catcttgtta caaggcatta aatctatcac ctctgattca aaggtatttg ccgtcgggag 16920
ctcaaaggtt gtttatagga gaaggttctg ggagcatgat gttattatat cagtctacat 16980
tggggcaatc aatttctttt tacaattcag gtatagatgg agattatata ccaggtcaaa 17040
gagaactgaa actatttccc tctgaatact caattgctga ggaagaccca tctctgacgg 17100
ggaaattgaa aggactagtg gtgcccctat tcaatggaag accagaaaca acatggatcg 17160
ggaatttaga ctcctacgag tatatcataa ataggacagc ggggcgaagt ataggtcttg 17220
tccattctga catggagtct gggattgaca aaaatgtaga ggagatacta gtagaacatt 17280
cccatctaat atctatcgcg ataaatgtta tgatggagga cggactatta gtatccaaga 17340
tagcatacac ccctggattc ccaatctcaa gattatttaa catgtacaga tcatatttcg 17400
gactagtact ggtgtgtttc ccagtatata gtaatccaga ttctactgaa gtatatcttc 17460
tttgcttaca gaagacggtc aagactattg ttcccccgca aaaagtcctt gagcactcta 17520
atttgcacga tgaagtcaat gaccagggaa taacatcagt gatttttaaa atcaagaatt 17580
cacagtctaa gcagttccac gatgatctaa agaagtacta tcagattgac caaccttttt 17640
ttgtaccaac taaaatcact agtgatgaac aagtacttct ccaagcaggg ctgaaactca 17700
atgggccaga aattcttaag agtgaaatca gttatgatat cggttcagat atcaatacat 17760
taagagacac catcataatt atgttaaatg aggctatgaa ttattttgat gacaacagat 17820
caccttcaca ccacctagaa ccctatccag ttttggagag aactagaatt aaaacaataa 17880
tgaattgtgt gactaaaaaa gtgattgtct actcacttat caagttcaag gacaccaaaa 17940
gctcagaact ttatcacatc aaaaataaca tcagaagaaa agttctaatc ttagatttca 18000
gatcgaagct catgacaaag actctaccta aagggatgca agagagaaga gaaaaaaacg 18060
gtttcaaaga agtttggata gtagatttat cgaatcgaga agttaaaatc tggtggaaga 18120
taatcggata catatctatt atctgattta accttccaaa tccaagacca actgataact 18180
tatgttgatc taaggttcag ttattaagaa aaacttaata acgattcttc tttacccttg 18240
ttcggt 18246
<210>9
<211>2244
<212>PRT
<213>Nipah virus
<400>9
Met Ala Asp Glu Leu Ser Ile Ser Asp Ile Ile Tyr Pro Glu Cys His
1 5 10 15
Leu Asp Ser Pro Ile Val Ser Gly Lys Leu Ile Ser Ala Ile Glu Tyr
20 25 30
Ala Gln Leu Arg His Asn Gln Pro Ser Asp Asp Lys Arg Leu Ser Glu
35 40 45
Asn Ile Arg Leu Asn Leu His Gly Lys Arg Lys Ser Leu Tyr Ile Leu
50 55 60
Arg Gln Ser Lys Gln Gly Asp Tyr Ile Arg Asn Asn Ile Lys Asn Leu
65 70 75 80
Lys Glu Phe Met His Ile Ala Tyr Pro Glu Cys Asn Asn Ile Leu Phe
85 90 95
Ser Ile Thr Ser Gln Gly Met Thr Ser Lys Leu Asp Asn Ile Met Lys
100 105 110
Lys Ser Phe Lys Ala Tyr Asn Ile Ile Ser Lys Lys Val Ile Gly Met
115 120 125
Leu Gln Asn Ile Thr Arg Asn Leu Ile Thr Gln Asp Arg Arg Asp Glu
130 135 140
Ile Ile Asn Ile His Glu Cys Arg Arg Leu Gly Asp Leu Gly Lys Asn
145 150 155 160
Met Ser Gln Ser Lys Trp Tyr Glu Cys Phe Leu Phe Trp Phe Thr Ile
165 170 175
Lys Thr Glu Met Arg Ala Val Ile Lys Asn Ser Gln Lys Pro Lys Phe
180 185 190
Arg Ser Asp Ser Cys Ile Ile His Met Arg Asp Lys Ser Thr Glu Ile
195 200 205
Ile Leu Asn Pro Asn Leu Ile Cys Ile Phe Lys Ser Asp Lys Thr Gly
210 215 220
Lys Lys Cys Tyr Tyr Leu Thr Pro Glu Met Val Leu Met Tyr Cys Asp
225 230 235 240
Val Leu Glu Gly Arg Met Met Met Glu Thr Thr Val Lys Ser Asp Ile
245 250 255
Lys Tyr Gln Pro Leu Ile Ser Arg Ser Asn Ala Leu Trp Gly Leu Ile
260 265 270
Asp Pro Leu Phe Pro Val Met Gly Asn Arg Ile Tyr Asn Ile Val Ser
275 280 285
Met Ile Glu Pro Leu Val Leu Ala Leu Leu Gln Leu Lys Asp Glu Ala
290 295 300
Arg Ile Leu Arg Gly Ala Phe Leu His His Cys Ile Lys Glu Met His
305 310 315 320
Gln Glu Leu Ser Glu Cys Gly Phe Thr Asp Gln Lys Ile Arg Ser Met
325 330 335
Phe Ile Asp Asp Leu Leu Ser Ile Leu Asn Ile Asp Asn Ile His Leu
340 345 350
Leu Ala Glu Phe Phe Ser Phe Phe Arg Thr Phe Gly His Pro Ile Leu
355 360 365
Glu Ala Lys Val Ala Ala Glu Lys Val Arg Glu His Met Leu Ala Asp
370 375 380
Lys Val Leu Glu Tyr Ala Pro Ile Met Lys Ala His Ala Ile Phe Cys
385 390 395 400
Gly Thr Ile Ile Asn Gly Tyr Arg Asp Arg His Gly Gly Ala Trp Pro
405 410 415
Pro Leu Tyr Leu Pro Ala His Ala Ser Lys His Ile Ile Arg Leu Lys
420 425 430
Asn Ser Gly Glu Ser Leu Thr Ile Asp Asp Cys Val Lys Asn Trp Glu
435 440 445
Ser Phe Cys Gly Ile Gln Phe Asp Cys Phe Met Glu Leu Lys Leu Asp
450 455 460
Ser Asp Leu Ser Met Tyr Met Lys Asp Lys Ala Leu Ser Pro Ile Lys
465 470 475 480
Asp Glu Trp Asp Ser Val Tyr Pro Arg Glu Val Leu Ser Tyr Thr Pro
485 490 495
Pro Lys Ser Thr Glu Pro Arg Arg Leu Val Asp Val Phe Val Asn Asp
500 505 510
Glu Asn Phe Asp Pro Tyr Asn Met Leu Glu Tyr Val Leu Ser Gly Ala
515 520 525
Tyr Leu Glu Asp Glu Gln Phe Asn Val Ser Tyr Ser Leu Lys Glu Lys
530 535 540
Glu Thr Lys Gln Ala Gly Arg Leu Phe Ala Lys Met Thr Tyr Lys Met
545 550 555 560
Arg Ala Cys Gln Val Ile Ala Glu Ala Leu Ile Ala Ser Gly Val Gly
565 570 575
Lys Tyr Phe Lys Glu Asn Gly Met Val Lys Asp Glu His Glu Leu Leu
580 585 590
Lys Thr Leu Phe Gln Leu Ser Ile Ser Ser Val Pro Arg Gly Asn Ser
595 600 605
Gln Gly Asn Asp Pro Gln Ser Ile Asn Asn Ile Glu Arg Asp Phe Gln
610 615 620
Tyr Phe Lys Gly Val Thr Thr Asn Val Lys Asp Lys Lys Asn Asn Ser
625 630 635 640
Phe Asn Lys Val Lys Ser Ala Leu Asn Asn Pro Cys Gln Ala Asp Gly
645 650 655
Val His His Asn Met Ser Pro Asn Thr Arg Asn Arg Tyr Lys Cys Ser
660 665 670
Asn Thr Ser Lys Ser Phe Leu Asp Tyr His Thr Glu Phe Asn Pro His
675 680 685
Asn His Tyr Lys Ser Asp Asn Thr Glu Ala Ala Val Leu Ser Arg Tyr
690 695 700
Glu Asp Asn Thr Gly Thr Lys Phe Asp Thr Val Ser Ala Phe Leu Thr
705 710 715 720
Thr Asp Leu Lys Lys Phe Cys Leu Asn Trp Arg Tyr Glu Ser Met Ala
725 730 735
Ile Phe Ala Glu Arg Leu Asp Glu Ile Tyr Gly Leu Pro Gly Phe Phe
740 745 750
Asn Trp Met His Lys Arg Leu Glu Arg Ser Val Ile Tyr Val Ala Asp
755 760 765
Pro Asn Cys Pro Pro Asn Ile Asp Lys His Met Glu Leu Glu Lys Thr
770 775 780
Pro Glu Asp Asp Ile Phe Ile His Tyr Pro Lys Gly Gly Ile Glu Gly
785 790 795 800
Tyr Ser Gln Lys Thr Trp Thr Ile Ala Thr Ile Pro Phe Leu Phe Leu
805 810 815
Ser Ala Tyr Glu Thr Asn Thr Arg Ile Ala Ala Ile Val Gln Gly Asp
820 825 830
Asn Glu Ser Ile Ala Ile Thr Gln Lys Val His Pro Asn Leu Pro Tyr
835 840 845
Lys Val Lys Lys Glu Ile Cys Ala Lys Gln Ala Gln Leu Tyr Phe Glu
850 855 860
Arg Leu Arg Met Asn Leu Arg Ala Leu Gly His Asn Leu Lys Ala Thr
865 870 875 880
Glu Thr Ile Ile Ser Thr His Leu Phe Ile Tyr Ser Lys Lys Ile His
885 890 895
Tyr Asp Gly Ala Val Leu Ser Gln Ala Leu Lys Ser Met Ser Arg Cys
900 905 910
Cys Phe Trp Ser Glu Thr Leu Val Asp Glu Thr Arg Ser Ala Cys Ser
915 920 925
Asn Ile Ser Thr Thr Ile Ala Lys Ala Ile Glu Asn Gly Leu Ser Arg
930 935 940
Asn Val Gly Tyr Cys Ile Asn Ile Leu Lys Val Ile Gln Gln Leu Leu
945 950 955 960
Ile Ser Thr Glu Phe Ser Ile Asn Glu Thr Leu Thr Leu Asp Val Thr
965 970 975
Ser Pro Ile Ser Asn Asn Leu Asp Trp Leu Ile Thr Ala Ala Leu Ile
980 985 990
Pro Ala Pro Ile Gly Gly Phe Asn Tyr Leu Asn Leu Ser Arg Ile Phe
995 1000 1005
Val Arg Asn Ile Gly Asp Pro Val Thr Ala Ser Leu Ala Asp Leu
1010 1015 1020
Lys Arg Met Ile Asp His Ser Ile Met Thr Glu Ser Val Leu Gln
1025 1030 1035
Lys Val Met Asn Gln Glu Pro Gly Asp Ala Ser Phe Leu Asp Trp
1040 1045 1050
Ala Ser Asp Pro Tyr Ser Gly Asn Leu Pro Asp Ser Gln Ser Ile
1055 1060 1065
Thr Lys Thr Ile Lys Asn Ile Thr Ala Arg Thr Ile Leu Arg Asn
1070 1075 1080
Ser Pro Asn Pro Met Leu Lys Gly Leu Phe His Asp Lys Ser Phe
1085 1090 1095
Asp Glu Asp Leu Glu Leu Ala Ser Phe Leu Met Asp Arg Arg Val
1100 1105 1110
Ile Leu Pro Arg Ala Ala His Glu Ile Leu Asp Asn Ser Leu Thr
1115 1120 1125
Gly Ala Arg Glu Glu Ile Ala Gly Leu Leu Asp Thr Thr Lys Gly
1130 1135 1140
Leu Ile Arg Ser Gly Leu Arg Lys Ser Gly Leu Gln Pro Lys Leu
1145 1150 1155
Val Ser Arg Leu Ser His His Asp Tyr Asn Gln Phe Leu Ile Leu
1160 1165 1170
Asn Lys Leu Leu Ser Asn Arg Arg Gln Asn Asp Leu Ile Ser Ser
1175 1180 1185
Asn Thr Cys Ser Val Asp Leu Ala Arg Ala Leu Arg Ser His Met
1190 1195 1200
Trp Arg Glu Leu Ala Leu Gly Arg Val Ile Tyr Gly Leu Glu Val
1205 1210 1215
Pro Asp Ala Leu Glu Ala Met Val Gly Arg Tyr Ile Thr Gly Ser
1220 1225 1230
Leu Glu Cys Gln Ile Cys Glu Gln Gly Asn Thr Met Tyr Gly Trp
1235 1240 1245
Phe Phe Val Pro Arg Asp Ser Gln Leu Asp Gln Val Asp Arg Glu
1250 1255 1260
His Ser Ser Ile Arg Val Pro Tyr Val Gly Ser Ser Thr Asp Glu
1265 1270 1275
Arg Ser Asp Ile Lys Leu Gly Asn Val Lys Arg Pro Thr Lys Ala
1280 1285 1290
Leu Arg Ser Ala Ile Arg Ile Ala Thr Val Tyr Thr Trp Ala Tyr
1295 1300 1305
Gly Asp Asn Glu Glu Cys Trp Tyr Glu Ala Trp Tyr Leu Ala Ser
1310 1315 1320
Gln Arg Val Asn Ile Asp Leu Asp Val Leu Lys Ala Ile Thr Pro
1325 1330 1335
Val Ser Thr Ser Asn Asn Leu Ser His Arg Leu Arg Asp Lys Ser
1340 1345 1350
Thr Gln Phe Lys Phe Ala Gly Ser Val Leu Asn Arg Val Ser Arg
1355 1360 1365
Tyr Val Asn Ile Ser Asn Asp Asn Leu Asp Phe Arg Ile Glu Gly
1370 1375 1380
Glu Lys Val Asp Thr Asn Leu Ile Tyr Gln Gln Ala Met Leu Leu
1385 1390 1395
Gly Leu Ser Val Leu Glu Gly Lys Phe Arg Leu Arg Leu Glu Thr
1400 1405 1410
Asp Asp Tyr Asn Gly Ile Tyr His Leu His Val Lys Asp Asn Cys
1415 1420 1425
Cys Val Lys Glu Val Ala Asp Val Gly Gln Val Asp Ala Glu Leu
1430 1435 1440
Pro Ile Pro Glu Tyr Thr Glu Val Asp Asn Asn His Leu Ile Tyr
1445 1450 1455
Asp Pro Asp Pro Val Ser Glu Ile Asp Cys Ser Arg Leu Ser Asn
1460 1465 1470
Gln Glu Ser Lys Ser Arg Glu Leu Asp Phe Pro Leu Trp Ser Thr
1475 1480 1485
Glu Glu Leu His Asp Val Leu Ala Lys Thr Val Ala Gln Thr Val
1490 1495 1500
Leu Glu Ile Ile Thr Lys Ala Asp Lys Asp Val Leu Lys Gln His
1505 1510 1515
Leu Ala Ile Asp Ser Asp Asp Asn Ile Asn Ser Leu Ile Thr Glu
1520 1525 1530
Phe Leu Ile Val Asp Pro Glu Leu Phe Ala Leu Tyr Leu Gly Gln
1535 1540 1545
Ser Ile Ser Ile Lys Trp Ala Phe Glu Ile His His Arg Arg Pro
1550 1555 1560
Arg Gly Arg His Thr Met Val Asp Leu Leu Ser Asp Leu Val Ser
1565 1570 1575
Asn Thr Ser Lys His Thr Tyr Lys Val Leu Ser Asn Ala Leu Ser
1580 1585 1590
His Pro Arg Val Phe Lys Arg Phe Val Asn Cys Gly Leu Leu Leu
1595 1600 1605
Pro Thr Gln Gly Pro Tyr Leu His Gln Gln Asp Phe Glu Lys Leu
1610 1615 1620
Ser Gln Asn Leu Leu Val Thr Ser Tyr Met Ile Tyr Leu Met Asn
1625 1630 1635
Trp Cys Asp Phe Lys Lys Ser Pro Phe Leu Ile Ala Glu Gln Asp
1640 1645 1650
Glu Thr Val Ile Ser Leu Arg Glu Asp Ile Ile Thr Ser Lys His
1655 1660 1665
Leu Cys Val Ile Ile Asp Leu Tyr Ala Asn His His Lys Pro Pro
1670 1675 1680
Trp Ile Ile Asp Leu Asn Pro Gln Glu Lys Ile Cys Val Leu Arg
1685 1690 1695
Asp Phe Ile Ser Lys Ser Arg His Val Asp Thr Ser Ser Arg Ser
1700 1705 1710
Trp Asn Thr Ser Asp Leu Asp Phe Val Ile Phe Tyr Ala Ser Leu
1715 1720 1725
Thr Tyr Leu Arg Arg Gly Ile Ile Lys Gln Leu Arg Ile Arg Gln
1730 1735 1740
Val Thr Glu Val Ile Asp Thr Thr Thr Met Leu Arg Asp Asn Ile
1745 1750 1755
Ile Val Glu Asn Pro Pro Ile Lys Thr Gly Val Leu Asp Ile Arg
1760 1765 1770
Gly Cys Ile Ile Tyr Asn Leu Glu Glu Ile Leu Ser Met Asn Thr
1775 1780 1785
Lys Ser Ala Ser Lys Lys Ile Phe Asn Leu Asn Ser Arg Pro Ser
1790 1795 1800
Val Glu Asn His Lys Tyr Arg Arg Ile Gly Leu Asn Ser Ser Ser
1805 1810 1815
Cys Tyr Lys Ala Leu Asn Leu Ser Pro Leu Ile Gln Arg Tyr Leu
1820 1825 1830
Pro Ser Gly Ala Gln Arg Leu Phe Ile Gly Glu Gly Ser Gly Ser
1835 1840 1845
Met Met Leu Leu Tyr Gln Ser Thr Leu Gly Gln Ser Ile Ser Phe
1850 1855 1860
Tyr Asn Ser Gly Ile Asp Gly Asp Tyr Ile Pro Gly Gln Arg Glu
1865 1870 1875
Leu Lys Leu Phe Pro Ser Glu Tyr Ser Ile Ala Glu Glu Asp Pro
1880 1885 1890
Ser Leu Thr Gly Lys Leu Lys Gly Leu Val Val Pro Leu Phe Asn
1895 1900 1905
Gly Arg Pro Glu Thr Thr Trp Ile Gly Asn Leu Asp Ser Tyr Glu
1910 1915 1920
Tyr Ile Ile Asn Arg Thr Ala Gly Arg Ser Ile Gly Leu Val His
1925 1930 1935
Ser Asp Met Glu Ser Gly Ile Asp Lys Asn Val Glu Glu Ile Leu
1940 1945 1950
Val Glu His Ser His Leu Ile Ser Ile Ala Ile Asn Val Met Met
1955 1960 1965
Glu Asp Gly Leu Leu Val Ser Lys Ile Ala Tyr Thr Pro Gly Phe
1970 1975 1980
Pro Ile Ser Arg Leu Phe Asn Met Tyr Arg Ser Tyr Phe Gly Leu
1985 1990 1995
Val Leu Val Cys Phe Pro Val Tyr Ser Asn Pro Asp Ser Thr Glu
2000 2005 2010
Val Tyr Leu Leu Cys Leu Gln Lys Thr Val Lys Thr Ile Val Pro
2015 2020 2025
Pro Gln Lys Val Leu Glu His Ser Asn Leu His Asp Glu Val Asn
2030 2035 2040
Asp Gln Gly Ile Thr Ser Val Ile Phe Lys Ile Lys Asn Ser Gln
2045 2050 2055
Ser Lys Gln Phe His Asp Asp Leu Lys Lys Tyr Tyr Gln Ile Asp
2060 2065 2070
Gln Pro Phe Phe Val Pro Thr Lys Ile Thr Ser Asp Glu Gln Val
2075 2080 2085
Leu Leu Gln Ala Gly Leu Lys Leu Asn Gly Pro Glu Ile Leu Lys
2090 2095 2100
Ser Glu Ile Ser Tyr Asp Ile Gly Ser Asp Ile Asn Thr Leu Arg
2105 2110 2115
Asp Thr Ile Ile Ile Met Leu Asn Glu Ala Met Asn Tyr Phe Asp
2120 2125 2130
Asp Asn Arg Ser Pro Ser His His Leu Glu Pro Tyr Pro Val Leu
2135 2140 2145
Glu Arg Thr Arg Ile Lys Thr Ile Met Asn Cys Val Thr Lys Lys
2150 2155 2160
Val Ile Val Tyr Ser Leu Ile Lys Phe Lys Asp Thr Lys Ser Ser
2165 2170 2175
Glu Leu Tyr His Ile Lys Asn Asn Ile Arg Arg Lys Val Leu Ile
2180 2185 2190
Leu Asp Phe Arg Ser Lys Leu Met Thr Lys Thr Leu Pro Lys Gly
2195 2200 2205
Met Gln Glu Arg Arg Glu Lys Asn Gly Phe Lys Glu Val Trp Ile
2210 2215 2220
Val Asp Leu Ser Asn Arg Glu Val Lys Ile Trp Trp Lys Ile Ile
2225 2230 2235
Gly Tyr Ile Ser Ile Ile
2240
<210>10
<211>602
<212>PRT
<213>Nipah virus
<400>10
Met Pro Ala Glu Asn Lys Lys Val Arg Phe Glu Asn Thr Thr Ser Asp
1 5 10 15
Lys Gly Lys Ile Pro Ser Lys Val Ile Lys Ser Tyr Tyr Gly Thr Met
20 25 30
Asp Ile Lys Lys Ile Asn Glu Gly Leu Leu Asp Ser Lys Ile Leu Ser
35 40 45
Ala Phe Asn Thr Val Ile Ala Leu Leu Gly Ser Ile Val Ile Ile Val
50 55 60
Met Asn Ile Met Ile Ile Gln Asn Tyr Thr Arg Ser Thr Asp Asn Gln
65 70 75 80
Ala Val Ile Lys Asp Ala Leu Gln Gly Ile Gln Gln Gln Ile Lys Gly
85 90 95
Leu Ala Asp Lys Ile Gly Thr Glu Ile Gly Pro Lys Val Ser Leu Ile
100 105 110
Asp Thr Ser Ser Thr Ile Thr Ile Pro Ala Asn Ile Gly Leu Leu Gly
115 120 125
Ser Lys Ile Ser Gln Ser Thr Ala Ser Ile Asn Glu Asn Val Asn Glu
130 135 140
Lys Cys Lys Phe Thr Leu Pro Pro Leu Lys Ile His Glu Cys Asn Ile
145 150 155 160
Ser Cys Pro Asn Pro Leu Pro Phe Arg Glu Tyr Arg Pro Gln Thr Glu
165 170 175
Gly Val Ser Asn Leu Val Gly Leu Pro Asn Asn Ile Cys Leu Gln Lys
180 185 190
Thr Ser Asn Gln Ile Leu Lys Pro Lys Leu Ile Ser Tyr Thr Leu Pro
195 200 205
Val Val Gly Gln Ser Gly Thr Cys Ile Thr Asp Pro Leu Leu Ala Met
210 215 220
Asp Glu Gly Tyr Phe Ala Tyr Ser His Leu Glu Arg Ile Gly Ser Cys
225 230 235 240
Ser Arg Gly Val Ser Lys Gln Arg Ile Ile Gly Val Gly Glu Val Leu
245 250 255
Asp Arg Gly Asp Glu Val Pro Ser Leu Phe Met Thr Asn Val Trp Thr
260 265 270
Pro Pro Asn Pro Asn Thr Val Tyr His Cys Ser Ala Val Tyr Asn Asn
275 280 285
Glu Phe Tyr Tyr Val Leu Cys Ala Val Ser Thr Val Gly Asp Pro Ile
290 295 300
Leu Asn Ser Thr Tyr Trp Ser Gly Ser Leu Met Met Thr Arg Leu Ala
305 310 315 320
Val Lys Pro Lys Ser Asn Gly Gly Gly Tyr Asn Gln His Gln Leu Ala
325 330 335
Leu Arg Ser Ile Glu Lys Gly Arg Tyr Asp Lys Val Met Pro Tyr Gly
340 345 350
Pro Ser Gly Ile Lys Gln Gly Asp Thr Leu Tyr Phe Pro Ala Val Gly
355 360 365
Phe Leu Val Arg Thr Glu Phe Lys Tyr Asn Asp Ser Asn Cys Pro Ile
370 375 380
Thr Lys Cys Gln Tyr Ser Lys Pro Glu Asn Cys Arg Leu Ser Met Gly
385 390 395 400
Ile Arg Pro Asn Ser His Tyr Ile Leu Arg Ser Gly Leu Leu Lys Tyr
405 410 415
Asn Leu Ser Asp Gly Glu Asn Pro Lys Val Val Phe Ile Glu Ile Ser
420 425 430
Asp Gln Arg Leu Ser Ile Gly Ser Pro Ser Lys Ile Tyr Asp Ser Leu
435 440 445
Gly Gln Pro Val Phe Tyr Gln Ala Ser Phe Ser Trp Asp Thr Met Ile
450 455 460
Lys Phe Gly Asp Val Leu Thr Val Asn Pro Leu Val Val Asn Trp Arg
465 470 475 480
Asn Asn Thr Val Ile Ser Arg Pro Gly Gln Ser Gln Cys Pro Arg Phe
485 490 495
Asn Thr Cys Pro Glu Ile Cys Trp Glu Gly Val Tyr Asn Asp Ala Phe
500 505 510
Leu Ile Asp Arg Ile Asn Trp Ile Ser Ala Gly Val Phe Leu Asp Ser
515 520 525
Asn Gln Thr Ala Glu Asn Pro Val Phe Thr Val Phe Lys Asp Asn Glu
530 535 540
Ile Leu Tyr Arg Ala Gln Leu Ala Ser Glu Asp Thr Asn Ala Gln Lys
545 550 555 560
Thr Ile Thr Asn Cys Phe Leu Leu Lys Asn Lys Ile Trp Cys Ile Ser
565 570 575
Leu Val Glu Ile Tyr Asp Thr Gly Asp Asn Val Ile Arg Pro Lys Leu
580 585 590
Phe Ala Val Lys Ile Pro Glu Gln Cys Thr
595 600
<210>11
<211>546
<212>PRT
<213>Nipah virus
<400>11
Met Val Val Ile Leu Asp Lys Arg Cys Tyr Cys Asn Leu Leu Ile Leu
1 5 10 15
Ile Leu Met Ile Ser Glu Cys Ser Val Gly Ile Leu His Tyr Glu Lys
20 25 30
Leu Ser Lys Ile Gly Leu Val Lys Gly Val Thr Arg Lys Tyr Lys Ile
35 40 45
Lys Ser Asn Pro Leu Thr Lys Asp Ile Val Ile Lys Met Ile Pro Asn
50 55 60
Val Ser Asn Met Ser Gln Cys Thr Gly Ser Val Met Glu Asn Tyr Lys
65 70 75 80
Thr Arg Leu Asn Gly Ile Lau Thr Pro Ile Lys Gly Ala Leu Glu Ile
85 90 95
Tyr Lys Asn Asn Thr His Asp Leu Val Gly Asp Val Arg Leu Ala Gly
100 105 110
Val Ile Met Ala Gly Val Ala Ile Gly Ile Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Val Ala Leu Tyr Glu Ala Met Lys Asn Ala Asp Asn Ile
130 135 140
Asn Lys Leu Lys Ser Ser Ile Glu Ser Thr Asn Glu Ala Val Val Lys
145 150 155 160
Leu Gln Glu Thr Ala Glu Lys Thr Val Tyr Val Leu Thr Ala Leu Gln
165 170 175
Asp Tyr Ile Asn Thr Asn Leu Val Pro Thr Ile Asp Lys Ile Ser Cys
180 185 190
Lys Gln Thr Glu Leu Ser Leu Asp Leu Ala Leu Ser Lys Tyr Leu Ser
195 200 205
Asp Leu Leu Phe Val Phe Gly Pro Asn Leu Gln Asp Pro Val Ser Asn
210 215 220
Ser Met Thr Ile Gln Ala Ile Ser Gln Ala Phe Gly Gly Asn Tyr Glu
225 230 235 240
Thr Leu Leu Arg Thr Leu Gly Tyr Ala Thr Glu Asp Phe Asp Asp Leu
245 250 255
Leu Glu Ser Asp Ser Ile Thr Gly Gln Ile Ile Tyr Val Asp Leu Ser
260 265 270
Ser Tyr Tyr Ile Ile Val Arg Val Tyr Phe Pro Ile Leu Thr Glu Ile
275 280 285
Gln Gln Ala Tyr Ile Gln Glu Leu Leu Pro Val Ser Phe Asn Asn Asp
290 295 300
Asn Ser Glu Trp Ile Ser Ile Val Pro Asn Phe Ile Leu Val Arg Asn
305 310 315 320
Thr Leu Ile Ser Asn Ile Glu Ile Gly Phe Cys Leu Ile Thr Lys Arg
325 330 335
Ser Val Ile Cys Asn Gln Asp Tyr Ala Thr Pro Met Thr Asn Asn Met
340 345 350
Arg Glu Cys Leu Thr Gly Ser Thr Glu Lys Cys Pro Arg Glu Leu Val
355 360 365
Val Ser Ser His Val Pro Arg Phe Ala Leu Ser Asn Gly Val Leu Phe
370 375 380
Ala Asn Cys Ile Ser Val Thr Cys Gln Cys Gln Thr Thr Gly Arg Ala
385 390 395 400
Ile Ser Gln Ser Gly Glu Gln Thr Leu Leu Met Ile Asp Asn Thr Thr
405 410 415
Cys Pro Thr Ala Val Leu Gly Asn Val Ile Ile Ser Leu Gly Lys Tyr
420 425 430
Leu Gly Ser Val Asn Tyr Asn Ser Glu Gly Ile Ala Ile Gly Pro Pro
435 440 445
Val Phe Thr Asp Lys Val Asp Ile Ser Ser Gln Ile Ser Ser Met Asn
450 455 460
Gln Ser Leu Gln Gln Ser Lys Asp Tyr Ile Lys Glu Ala Gln Arg Leu
465 470 475 480
Leu Asp Thr Val Asn Pro Ser Leu Ile Ser Met Leu Ser Met Ile Ile
485 490 495
Leu Tyr Val Leu Ser Ile Ala Ser Leu Cys Ile Gly Leu Ile Thr Phe
500 505 510
Ile Ser Phe Ile Ile Val Glu Lys Lys Arg Asn Thr Tyr Ser Arg Leu
515 520 525
Glu Asp Arg Arg Val Arg Pro Thr Ser Ser Gly Asp Leu Tyr Tyr Ile
530 535 540
Gly Thr
545
<210>12
<211>352
<212>PRT
<213>Nipah virus
<400>12
Met Glu Pro Asp Ile Lys Ser Ile Ser Ser Glu Ser Met Glu Gly Val
1 5 10 15
Ser Asp Phe Ser Pro Ser Ser Trp Glu His Gly Gly Tyr Leu Asp Lys
20 25 30
Val Glu Pro Glu Ile Asp Glu Asn Gly Ser Met Ile Pro Lys Tyr Lys
35 40 45
Ile Tyr Thr Pro Gly Ala Asn Glu Arg Lys Tyr Asn Asn Tyr Met Tyr
50 55 60
Leu Ile Cys Tyr Gly Phe Val Glu Asp Val Glu Arg Thr Pro Glu Thr
65 70 75 80
Gly Lys Arg Lys Lys Ile Arg Thr Ile Ala Ala Tyr Pro Leu Gly Val
85 90 95
Gly Lys Ser Ala Ser His Pro Gln Asp Leu Leu Glu Glu Leu Cys Ser
100 105 110
Leu Lys Val Thr Val Arg Arg Thr Ala Gly Ser Thr Glu Lys Ile Val
115 120 125
Phe Gly Ser Ser Gly Pro Leu Asn His Leu Val Pro Trp Lys Lys Val
130 135 140
Leu Thr Ser Gly Ser Ile Phe Asn Ala Val Lys Val Cys Arg Asn Val
145 150 155 160
Asp Gln Ile Gln Leu Asp Lys His Gln Ala Leu Arg Ile Phe Phe Leu
165 170 175
Ser Ile Thr Lys Leu Asn Asp Ser Gly Ile Tyr Met Ile Pro Arg Thr
180 185 190
Met Leu Glu Phe Arg Arg Asn Asn Ala Ile Ala Phe Asn Leu Leu Val
195 200 205
Tyr Leu Lys Ile Asp Ala Asp Leu Ser Lys Met Gly Ile Gln Gly Ser
210 215 220
Leu Asp Lys Asp Gly Phe Lys Val Ala Ser Phe Met Leu His Leu Gly
225 230 235 240
Asn Phe Val Arg Arg Ala Gly Lys Tyr Tyr Ser Val Asp Tyr Cys Arg
245 250 255
Arg Lys Ile Asp Arg Met Lys Leu Gln Phe Ser Leu Gly Ser Ile Gly
260 265 270
Gly Leu Ser Leu His Ile Lys Ile Asn Gly Val Ile Ser Lys Arg Leu
275 280 285
Phe Ala Gln Met Gly Phe Gln Lys Asn Leu Cys Phe Ser Leu Met Asp
290 295 300
Ile Asn Pro Trp Leu Asn Arg Leu Thr Trp Asn Asn Ser Cys Glu Ile
305 310 315 320
Ser Arg Val Ala Ala Val Leu Gln Pro Ser Ile Pro Arg Glu Phe Met
325 330 335
Ile Tyr Asp Asp Val Phe Ile Asp Asn Thr Gly Arg Ile Leu Lys Gly
340 345 350
<210>13
<211>166
<212>PRT
<213>Nipah virus
<400>13
Met Met Ala Ser Ile Leu Leu Thr Leu Phe Arg Arg Thr Lys Lys Lys
1 5 10 15
Tyr Arg Arg His Thr Asp Asp Gln Val Phe Asn Asn Pro Ala Ser Lys
20 25 30
Ile Lys Gln Lys Pro Gly Lys Ile Phe Cys Ser Ala Pro Val Glu Asn
35 40 45
Leu Asn Lys Leu Arg Gly Glu Cys Leu Arg Met Met Glu Met Leu Lys
50 55 60
Glu Glu Thr Trp Arg Ile Tyr Pro Val Leu Leu Pro Gln Met Glu Leu
65 70 75 80
Leu Glu Arg Glu Cys Arg Thr Pro Val Thr Gly Gln Lys Val Gln Met
85 90 95
Thr Tyr Asn Trp Thr Gln Trp Leu Gln Thr Leu Tyr Thr Met Ile Met
100 105 110
Glu Glu Asn Val Pro Asp Met Asp Leu Leu Gln Ala Leu Arg Glu Gly
115 120 125
Gly Val Ile Thr His Gln Glu Gln Thr Met Gly Met Tyr Val Leu Tyr
130 135 140
Leu Met Gln Arg Cys Cys Pro Met Leu Pro Lys Leu Gln Phe Leu Lys
145 150 155 160
Lys Ile Gly Lys Leu Ile
165
<210>14
<211>456
<212>PRT
<213>Nipah virus
<400>14
Met Asp Lys Leu Glu Leu Val Asn Asp Gly Leu Asn Ile Ile Asp Phe
1 5 10 15
Ile Gln Lys Asn Gln Lys Glu Ile Gln Lys Thr Tyr Gly Arg Ser Ser
20 25 30
Ile Gln Gln Pro Ser Ile Lys Asp Gln Thr Lys Ala Trp Glu Asp Phe
35 40 45
Leu Gln Cys Thr Ser Gly Glu Ser Glu Gln Val Glu Gly Gly Met Ser
50 55 60
Lys Asp Asp Gly Asp Val Glu Arg Arg Asn Leu Glu Asp Leu Ser Ser
65 70 75 80
Thr Ser Pro Thr Asp Gly Thr Ile Gly Lys Arg Val Ser Asn Thr Arg
85 90 95
Asp Trp Ala Glu Gly Ser Asp Asp Ile Gln Leu Asp Pro Val Val Thr
100 105 110
Asp Val Val Tyr His Asp His Gly Gly Glu Cys Thr Gly Tyr Gly Phe
115 120 125
Thr Ser Ser Pro Glu Arg Gly Trp Ser Asp Tyr Thr Ser Gly Ala Asn
130 135 140
Asn Gly Asn Val Cys Leu Val Ser Asp Ala Lys Met Leu Ser Tyr Ala
145 150 155 160
Pro Glu Ile Ala Val Ser Lys Glu Asp Arg Glu Thr Asp Leu Val His
165 170 175
Leu Glu Asn Lys Leu Ser Thr Thr Gly Leu Asn Pro Thr Ala Val Pro
180 185 190
Phe Thr Leu Arg Asn Leu Ser Asp Pro Ala Lys Asp Ser Pro Val Ile
195 200 205
Ala Glu His Tyr Tyr Gly Leu Gly Val Lys Glu Gln Asn Val Gly Pro
210 215 220
Gln Thr Ser Arg Asn Val Asn Leu Asp Ser Ile Lys Leu Tyr Thr Ser
225 230 235 240
Asp Asp Glu Glu Ala Asp Gln Leu Glu Phe Glu Asp Glu Phe Ala Gly
245 250 255
Ser Ser Ser Glu Val Ile Val Gly Ile Ser Pro Glu Asp Glu Glu Pro
260 265 270
Ser Ser Val Gly Gly Lys Pro Asn Glu Ser Ile Gly Arg Thr Ile Glu
275 280 285
Gly Gln Ser Ile Arg Asp Asn Leu Gln Ala Lys Asp Asn Lys Ser Thr
290 295 300
Asp Val Pro Gly Ala Gly Pro Lys Asp Ser Ala Val Lys Glu Glu Pro
305 310 315 320
Pro Gln Lys Arg Leu Pro Met Leu Ala Glu Glu Phe Glu Cys Ser Gly
325 330 335
Ser Glu Asp Pro Ile Ile Arg Glu Leu Leu Lys Glu Asn Ser Leu Ile
340 345 350
Asn Cys Gln Gln Gly Lys Asp Ala Gln Pro Pro Tyr His Trp Ser Ile
355 360 365
Glu Arg Ser Ile Ser Pro Asp Lys Thr Glu Ile Val Asn Gly Ala Val
370 375 380
Gln Thr Ala Asp Arg Gln Arg Pro Gly Thr Pro Met Pro Lys Ser Arg
385 390 395 400
Gly Ile Pro Ile Lys Lys Gly His Arg Arg Glu Ile Ser Ile Cys Trp
405 410 415
Asp Gly Lys Arg Ala Trp Val Glu Glu Trp Cys Asn Pro Ala Cys Ser
420 425 430
Arg Ile Thr Pro Leu Pro Arg Arg Gln Glu Cys Gln Cys Gly Glu Cys
435 440 445
Pro Thr Glu Cys Phe His Cys Gly
450 455
<210>15
<211>709
<212>PRT
<213>Nipah virus
<400>15
Met Asp Lys Leu Glu Leu Val Asn Asp Gly Leu Asn Ile Ile Asp Phe
1 5 10 15
Ile Gln Lys Asn Gln Lys Glu Ile Gln Lys Thr Tyr Gly Arg Ser Ser
20 25 30
Ile Gln Gln Pro Ser Ile Lys Asp Gln Thr Lys Ala Trp Glu Asp Phe
35 40 45
Leu Gln Cys Thr Ser Gly Glu Ser Glu Gln Val Glu Gly Gly Met Ser
50 55 60
Lys Asp Asp Gly Asp Val Glu Arg Arg Asn Leu Glu Asp Leu Ser Ser
65 70 75 80
Thr Ser Pro Thr Asp Gly Thr Ile Gly Lys Arg Val Ser Asn Thr Arg
85 90 95
Asp Trp Ala Glu Gly Ser Asp Asp Ile Gln Leu Asp Pro Val Val Thr
100 105 110
Asp Val Val Tyr His Asp His Gly Gly Glu Cys Thr Gly Tyr Gly Phe
115 120 125
Thr Ser Ser Pro Glu Arg Gly Trp Ser Asp Tyr Thr Ser Gly Ala Asn
130 135 140
Asn Gly Asn Val Cys Leu Val Ser Asp Ala Lys Met Leu Ser Tyr Ala
145 150 155 160
Pro Glu Ile Ala Val Ser Lys Glu Asp Arg Glu Thr Asp Leu Val His
165 170 175
Leu Glu Asn Lys Leu Ser Thr Thr Gly Leu Asn Pro Thr Ala Val Pro
180 185 190
Phe Thr Leu Arg Asn Leu Ser Asp Pro Ala Lys Asp Ser Pro Val Ile
195 200 205
Ala Glu His Tyr Tyr Gly Leu Gly Val Lys Glu Gln Asn Val Gly Pro
210 215 220
Gln Thr Ser Arg Asn Val Asn Leu Asp Ser Ile Lys Leu Tyr Thr Ser
225 230 235 240
Asp Asp Glu Glu Ala Asp Gln Leu Glu Phe Glu Asp Glu Phe Ala Gly
245 250 255
Ser Ser Ser Glu Val Ile Val Gly Ile Ser Pro Glu Asp Glu Glu Pro
260 265 270
Ser Ser Val Gly Gly Lys Pro Asn Glu Ser Ile Gly Arg Thr Ile Glu
275 280 285
Gly Gln Ser Ile Arg Asp Asn Leu Gln Ala Lys Asp Asn Lys Ser Thr
290 295 300
Asp Val Pro Gly Ala Gly Pro Lys Asp Ser Ala Val Lys Glu Glu Pro
305 310 315 320
Pro Gln Lys Arg Leu Pro Met Leu Ala Glu Glu Phe Glu Cys Ser Gly
325 330 335
Ser Glu Asp Pro Ile Ile Arg Glu Leu Leu Lys Glu Asn Ser Leu Ile
340 345 350
Asn Cys Gln Gln Gly Lys Asp Ala Gln Pro Pro Tyr His Trp Ser Ile
355 360 365
Glu Arg Ser Ile Ser Pro Asp Lys Thr Glu Ile Val Asn Gly Ala Val
370 375 380
Gln Thr Ala Asp Arg Gln Arg Pro Gly Thr Pro Met Pro Lys Ser Arg
385 390 395 400
Gly Ile Pro Ile Lys Lys Gly Thr Asp Ala Lys Tyr Pro Ser Ala Gly
405 410 415
Thr Glu Asn Val Pro Gly Ser Lys Ser Gly Ala Thr Arg His Val Arg
420 425 430
Gly Ser Pro Pro Tyr Gln Glu Gly Lys Ser Val Asn Ala Glu Asn Val
435 440 445
Gln Leu Asn Ala Ser Thr Ala Val Lys Glu Thr Asp Lys Ser Glu Val
450 455 460
Asn Pro Val Asp Asp Asn Asp Ser Leu Asp Asp Lys Tyr Ile Met Pro
465 470 475 480
Ser Asp Asp Phe Ser Asn Thr Phe Phe Pro His Asp Thr Asp Arg Leu
485 490 495
Asn Tyr His Ala Asp His Leu Gly Asp Tyr Asp Leu Glu Thr Leu Cys
500 505 510
Glu Glu Ser Val Leu Met Gly Val Ile Asn Ser Ile Lys Leu Ile Asn
515 520 525
Leu Asp Met Arg Leu Asn His Ile Glu Glu Gln Val Lys Glu Ile Pro
530 535 540
Lys Ile Ile Asn Lys Leu Glu Ser Ile Asp Arg Val Leu Ala Lys Thr
545 550 555 560
Asn Thr Ala Leu Ser Thr Ile Glu Gly His Leu Val Ser Met Met Ile
565 570 575
Met Ile Pro Gly Lys Gly Lys Gly Glu Arg Lys Gly Lys Asn Asn Pro
580 585 590
Glu Leu Lys Pro Val Ile Gly Arg Asp Ile Leu Glu Gln Gln Ser Leu
595 600 605
Phe Ser Phe Asp Asn Val Lys Asn Phe Arg Asp Gly Ser Leu Thr Asn
610 615 620
Glu Pro Tyr Gly Ala Ala Val Gln Leu Arg Glu Asp Leu Ile Leu Pro
625 630 635 640
Glu Leu Asn Phe Glu Glu Thr Asn Ala Ser Gln Phe Val Pro Met Ala
645 650 655
Asp Asp Ser Ser Arg Asp Val Ile Lys Thr Leu Ile Arg Thr His Ile
660 665 670
Lys Asp Arg Glu Leu Arg Ser Glu Leu Ile Gly Tyr Leu Asn Lys Ala
675 680 685
Glu Asn Asp Glu Glu Ile Gln Glu Ile Ala Asn Thr Val Asn Asp Ile
690 695 700
Ile Asp Gly Asn Ile
705
<210>16
<211>532
<212>PRT
<213>Nipah virus
<400>16
Met Ser Asp Ile Phe Glu Glu Ala Ala Ser Phe Arg Ser Tyr Gln Ser
1 5 10 15
Lys Leu Gly Arg Asp Gly Arg Ala Ser Ala Ala Thr Ala Thr Leu Thr
20 25 30
Thr Lys Ile Arg Ile Phe Val Pro Ala Thr Asn Ser Pro Glu Leu Arg
35 40 45
Trp Glu Leu Thr Leu Phe Ala Leu Asp Val Ile Arg Ser Pro Ser Ala
50 55 60
Ala Glu Ser Met Lys Val Gly Ala Ala Phe Thr Leu Ile Ser Met Tyr
65 70 75 80
Ser Glu Arg Pro Gly Ala Leu Ile Arg Ser Leu Leu Asn Asp Pro Asp
85 90 95
Ile Glu Ala Val Ile Ile Asp Val Gly Ser Met Val Asn Gly Ile Pro
100 105 110
Val Met Glu Arg Arg Gly Asp Lys Ala Gln Glu Glu Met Glu Gly Leu
115 120 125
Met Arg Ile Leu Lys Thr Ala Arg Asp Ser Ser Lys Gly Lys Thr Pro
130 135 140
Phe Val Asp Ser Arg Ala Tyr Gly Leu Arg Ile Thr Asp Met Ser Thr
145 150 155 160
Leu Val Ser Ala Val Ile Thr Ile Glu Ala Gln Ile Trp Ile Leu Ile
165 170 175
Ala Lys Ala Val Thr Ala Pro Asp Thr Ala Glu Glu Ser Glu Thr Arg
180 185 190
Arg Trp Ala Lys Tyr Val Gln Gln Lys Arg Val Asn Pro Phe Phe Ala
195 200 205
Leu Thr Gln Gln Trp Leu Thr Glu Met Arg Asn Leu Leu Ser Gln Ser
210 215 220
Leu Ser Val Arg Lys Phe Met Val Glu Ile Leu Ile Glu Val Lys Lys
225 230 235 240
Gly Gly Ser Ala Lys Gly Arg Ala Val Glu Ile Ile Ser Asp Ile Gly
245 250 255
Asn Tyr Val Glu Glu Thr Gly Met Ala Gly Phe Phe Ala Thr Ile Arg
260 265 270
Phe Gly Leu Glu Thr Arg Tyr Pro Ala Leu Ala Leu Asn Glu Phe Gln
275 280 285
Ser Asp Leu Asn Thr Ile Lys Ser Leu Met Leu Leu Tyr Arg Glu Ile
290 295 300
Gly Pro Arg Ala Pro Tyr Met Val Leu Leu Glu Glu Ser Ile Gln Thr
305 310 315 320
Lys Phe Ala Pro Gly Gly Tyr Pro Leu Leu Trp Ser Phe Ala Met Gly
325 330 335
Val Ala Thr Thr Ile Asp Arg Ser Met Gly Ala Leu Asn Ile Asn Arg
340 345 350
Gly Tyr Leu Glu Pro Met Tyr Phe Arg Leu Gly Gln Lys Ser Ala Arg
355 360 365
His His Ala Gly Gly Ile Asp Gln Asn Met Ala Asn Arg Leu Gly Leu
370 375 380
Ser Ser Asp Gln Val Ala Glu Leu Ala Ala Ala Val Gln Glu Thr Ser
385 390 395 400
Ala Gly Arg Gln Glu Ser Asn Val Gln Ala Arg Glu Ala Lys Phe Ala
405 410 415
Ala Gly Gly Val Leu Ile Gly Gly Ser Asp Gln Asp Ile Asp Glu Gly
420 425 430
Glu Glu Pro Ile Glu Gln Ser Gly Arg Gln Ser Val Thr Phe Lys Arg
435 440 445
Glu Met Ser Ile Ser Ser Leu Ala Asn Ser Val Pro Ser Ser Ser Val
450 455 460
Ser Thr Ser Gly Gly Thr Arg Leu Thr Asn Ser Leu Leu Asn Leu Arg
465 470 475 480
Ser Arg Leu Ala Ala Lys Ala Ala Lys Glu Ala Ala Ser Ser Asn Ala
485 490 495
Thr Asp Asp Pro Ala Ile Ser Asn Arg Thr Gln Gly Glu Ser Glu Lys
500 505 510
Lys Asn Asn Gln Asp Leu Lys Pro Ala Gln Asn Asp Leu Asp Phe Val
515 520 525
Arg Ala Asp Val
530
<210>17
<211>18246
<212>DNA
<213>Nipah virus
<400>17
accaaacaag ggagaatatg gatacgttaa aatatataac gtatttttaa aacttaggaa 60
ccaagacaaa cacttttggt cttggtattg gatcctcaag aaatatatca tcatgagtga 120
tatctttgaa gaggcggcta gttttaggag ttatcaatct aagttaggga gagatgggag 180
ggctagtgca gcaactgcta ctttgacaac caagataagg atatttgtac cagctactaa 240
tagtccagag ctcagatggg aactaacatt gtttgcactt gatgtgatta gatctccgag 300
tgctgccgag tcaatgaaag ttggagctgc tttcacactc atctctatgt attcagagag 360
acccggggct ctcattagaa gtctcctcaa tgacccagac attgaagctg taataataga 420
tgttggatca atggtcaacg gaataccagt aatggagagg agaggagaca aggctcagga 480
ggagatggaa ggcttgatga gaatcctcaa aactgctcga gacagcagca agggaaaaac 540
accttttgtt gacagccgag cttacggcct acggataaca gacatgagca ccctggtctc 600
tgcagttatc accatcgagg cccagatctg gatactgatc gctaaagcag ttacagctcc 660
cgacactgcc gaggaaagtg aaactagaag atgggctaaa tacgtccaac aaaagagagt 720
caatccgttc tttgctctaa ctcagcaatg gctaacagaa atgaggaatc tgctctccca 780
gagtctatca gtaaggaagt tcatggttga gatcctcata gaagtcaaga aaggaggatc 840
tgctaaaggc agagcagtag aaataatctc agacatcgga aactatgtcg aggaaactgg 900
tatggcagga ttcttcgcaa ccatcagatt cgggttggag acaaggtatc cagcacttgc 960
actcaacgaa ttccagagtg acctcaacac catcaaaagc ttgatgctac tctacagaga 1020
aattggccca agagcccctt atatggtgct tcttgaagaa tcaattcaga ctaaatttgc 1080
ccctggaggt tacccattat tgtggagctt tgccatgggt gtggctacta ctattgacag 1140
gtctatgggg gcattgaata tcaatcgtgg ttatcttgag cctatgtatt tcagactagg 1200
ccaaaaatca gcacgtcacc atgctggagg aattgatcag aacatggcaa atagactggg 1260
actaagttca gatcaagttg cagaactcgc tgctgcagtt caggaaacat cagcaggaag 1320
gcaagagagt aatgttcagg ctagagaggc aaaatttgct gcaggaggtg tgctcattgg 1380
aggcagtgat caagatatcg atgaagggga agaacctata gaacagagtg gcagacagtc 1440
agttaccttc aaaagggaga tgagtatttc atcccttgct aacagtgtgc cgagcagttc 1500
tgtgagcaca tccggtggga ccagattgac taattcatta ctaaacctca gatcaagact 1560
ggctgcaaaa gcagcaaaag aagccgcctc atccaatgca acagatgatc cagcaatcag 1620
caacagaact caaggggaat cagagaagaa gaataatcaa gacctcaaac ctgctcaaaa 1680
tgaccttgat ttcgtcagag ctgatgtgtg acgtctattt ccaatattct acagtatcca 1740
aaaatctttc tatagtacac tatcataata cgacactaag ggatcaacca tatcaaagtt 1800
acgaatcgtt ttaattatat taatcaaatg atactctttt atgggcaaac cgaagaacca 1860
atgtctacat gtaaattgag ctttggtatt gcaatctaat acttgctcaa aatcttgaac 1920
tattagtgta atttctatca tcatagagtt atcaagattt tattatataa gttggtgcag 1980
atctttggac atgaattaca cactacactc taatgaagac aaaatttaca ttacatattt 2040
aaggactatt tcctatcctt tcaatggtac ttggttatga aggtttctta atttaactaa 2100
gctactgtct ttgcactgga atatacaata cctcttacct catttcttac tttaatatca 2160
tgttattttt ttgataagtc acttaacttg accaaggtct accaggtaat gctcgcacaa 2220
gtgaactgca atctcaactt agattaaaca taatcatgca aaatcactat tttgtactac 2280
taactcatta agaaaaactt aggatccaag agatttactc taggatctcc tattaagctt 2340
agcagtcatt agttgagagt tcaacttgca aaactctaac cttcactcta ataacaattc 2400
atccaatgga taaattggaa ctagtcaatg atggcctcaa tattattgac tttattcaga 2460
agaaccaaaa agaaatacag aagacatacg gacgatcaag tattcaacaa cccagcatca 2520
aagatcaaac aaaagcctgg gaagattttc tgcagtgcac cagtggagaa tctgaacaag 2580
ttgagggggg aatgtctaag gatgatggag atgttgaaag aagaaacttg gaggatctat 2640
ccagtacttc tcccacagat ggaactattg gaaagagagt gtcgaacacc cgtgactggg 2700
cagaaggttc agatgacata caactggacc cagtggttac agacgttgta taccatgatc 2760
atggaggaga atgtaccgga tatggattta cttcaagccc tgagagaggg tggagtgatt 2820
acacatcagg agcaaacaat gggaatgtat gtcttgtatc tgatgcaaag atgctgtcct 2880
atgctcccga aattgcagtt tctaaagaag atcgggaaac tgatctagtt catcttgaga 2940
ataaactatc tactacagga ctgaatccca cagcagtacc gttcactctg agaaacctgt 3000
ctgatcctgc aaaagactct cctgtgattg ctgaacacta ctacggacta ggagttaaag 3060
agcaaaacgt tggccctcag actagcagaa atgtcaattt ggacagcatc aaattgtaca 3120
catcagatga cgaagaggca gatcagcttg aattcgaaga tgagtttgca ggaagctcaa 3180
gtgaagtgat agtcggcatt tctcctgaag atgaagagcc ttcaagtgtt ggcggaaaac 3240
ccaatgaatc cattggacgt acaatcgaag gccaatcaat ccgagacaac cttcaagcca 3300
aggacaacaa atcaacagat gtaccaggag caggaccgaa agattcagca gtgaaggaag 3360
aaccacccca gaagaggcta cctatgttag ctgaagaatt tgagtgctct ggatcggaag 3420
acccaatcat tcgggagctg ctgaaggaga actcactcat aaattgtcag caagggaaag 3480
atgctcagcc tccatatcat tggagcatcg agaggtcaat aagcccggat aaaactgaga 3540
tcgtcaacgg tgctgtgcaa actgctgaca ggcaaagacc aggaactccg atgccaaagt 3600
cccgaggtat tcccattaaa aagggcacag acgcgaaata tccatctgct gggacggaaa 3660
acgtgcctgg gtcgaagagt ggtgcaaccc ggcatgttcg aggatcaccc ccctaccaag 3720
aaggcaagag tgtcaatgcg gagaatgtcc aactgaatgc ttccactgcg gttaaggaaa 3780
ctgataagtc agaagtaaac cccgtagacg acaacgactc acttgatgat aaatacatca 3840
tgccttcaga tgatttctca aacactttct tcccgcacga cactgatcgc ttgaattatc 3900
acgcagatca tttaggtgat tatgaccttg aaaccctgtg tgaagagtcg gttctaatgg 3960
gagtgatcaa ctctataaaa ttaattaatc tggatatgcg cttaaatcac attgaagaac 4020
aagttaaaga gatcccaaag atcatcaata agcttgagtc cattgacaga gttctggcca 4080
agactaacac cgcactctca accattgaag gacacctggt ttccatgatg ataatgatac 4140
cagggaaagg gaaaggagaa agaaagggga aaaataatcc tgagcttaaa ccagtgatag 4200
gaagagacat tctagagcag caatctcttt tttcttttga caatgtcaag aatttcagag 4260
atggatcgtt gacaaacgaa ccgtatgggg cagctgtaca gttgagagaa gatcttattc 4320
ttcctgaact taattttgag gagacaaatg catctcaatt tgttcctatg gcagatgatt 4380
catccagaga tgttatcaag acattgataa ggactcacat taaagataga gagttgagat 4440
cagaactgat tggttacctg aataaagcgg aaaatgatga ggaaattcag gagatagcga 4500
acactgtcaa tgacatcatt gacggtaata tttgatcact gaattgtcag cagaaataca 4560
atgatctaac aacaatctcc cacaagtaga caatggtttc aggtcaataa taacaacctc 4620
aatactaatc tttcacataa gcattactca ttccagccct cagacgataa cacaatactt 4680
gatacatgtt tattgaagtg tatgtagcat gattgaacta ttcaataact gtatttctca 4740
ctcttgctct tagttagtca ttgtgtctaa taattattat tacagtacaa ggtattatga 4800
attcaaagat acgcaataaa tctgatatca gcatagagta gaaaattgtt gtttttgtca 4860
tgatcattcg aagatttaac aatgatgtca actttcatac ctaaacataa taacataaaa 4920
tggtcgattt gtattgtaga tctctcacgc attttagtgt catgaattag tgtttcaaat 4980
cagttgcata tcaattaaga aaaacttagg agacaggtat agaacctctc tttcagataa 5040
ctggtcaatt aaggacagaa attctgtttc tcaaatccgc tagcctttgt caaagaggac 5100
acaagcaatg gagccggaca tcaagagtat ttcaagtgag tcaatggaag gagtatctga 5160
tttcagccct agttcttggg agcatggtgg gtatcttgat aaggttgaac cagaaattga 5220
tgaaaatggc agtatgattc caaaatacaa gatctatacc ccaggagcta acgagaggaa 5280
atacaacaac tacatgtacc ttatatgtta cggctttgtt gaagatgttg agagaacccc 5340
agagacaggg aaacgcaaga agatcaggac aattgctgcc taccctctgg gtgttggtaa 5400
gagtgcctct catccccaag atcttctgga ggaactctgt tccctcaaag ttactgtgag 5460
aagaacagct ggatcaactg agaaaattgt gtttggatca tctggccctc taaatcacct 5520
cgttccgtgg aagaaagtac tgactagtgg ttcaattttt aatgcagtca aggtttgtcg 5580
gaacgttgat cagatacagc ttgacaagca tcaagctctg agaatatttt ttctcagtat 5640
cacaaagctc aatgattctg gaatctacat gattccacga accatgcttg agttcaggag 5700
aaacaatgcc attgccttca atcttctagt gtacttgaag attgatgctg atttatccaa 5760
aatggggatc cagggaagcc tcgataaaga tggcttcaag gttgcctcct tcatgctaca 5820
cttggggaac tttgtccgtc gtgcagggaa gtattactct gttgattatt gtaggaggaa 5880
gattgatagg atgaaattgc agttttcact gggttccata ggcggactaa gtctccacat 5940
taagatcaat ggtgtaatca gcaaacggct gtttgctcaa atgggattcc aaaaaaacct 6000
ttgtttctct ttgatggaca tcaatccttg gctcaacaga ttgacctgga acaacagttg 6060
tgagatcagc cgagtagcag ctgtgttgca gccttctatt ccaagagagt tcatgatcta 6120
tgatgatgtc ttcattgaca atacagggag aattctaaag ggctaaacag aattcttcta 6180
aaatttaatc agtcatgagt ttagtaatca tacctagtca taatacatca cacaggacta 6240
tttacaaaag acagttaaaa aatggaataa tcatgtagta gtaattgaga acattattag 6300
aatagtataa ctaaaatgta gtttttttga gtatttgatt taaaattaga taactattac 6360
aaaaaactta ggagccaagc tcttgcctcg ttcagaaggt taaacaagca ttcttaccat 6420
tggatcaaca aaaggattgg ttttatcgtc taagaaattt attgaaaggc aaagaaattc 6480
ctggttttat gttgaatgag gtgtatcaaa ctaaggagac cttctaacag ccaggtcata 6540
ggaatataaa taaaaataag aataaaattg attccatcgg aagattcatt tcaagaagtg 6600
atcaaatcaa agcggttggc agacctacca atcatatacc acaagactcg acaatggtag 6660
ttatacttga caagagatgt tattgtaatc ttttaatatt gattttgatg atctcggagt 6720
gtagtgttgg gattctacat tatgagaaat tgagtaaaat tggacttgtc aaaggagtaa 6780
caagaaaata caagattaaa agcaatcctc tcacaaaaga cattgttata aaaatgattc 6840
cgaatgtgtc gaacatgtct cagtgcacag ggagtgtcat ggaaaattat aaaacacgat 6900
taaacggtat cttaacacct ataaagggag cgttagagat ctacaaaaac aacactcatg 6960
accttgtcgg tgatgtgaga ttagccggag ttataatggc aggagttgct attgggattg 7020
caaccgcagc tcaaatcact gcaggtgtag cactatatga ggcaatgaag aatgctgaca 7080
acatcaacaa actcaaaagc agcattgaat caactaatga agctgtcgtt aaacttcaag 7140
agactgcaga aaagacagtc tatgtgctga ctgctctaca ggattacatt aatactaatt 7200
tagtaccgac aattgacaag ataagctgca aacagacaga actctcacta gatctggcat 7260
tatcaaagta cctctctgat ttgctttttg tatttggccc caaccttcaa gacccagttt 7320
ctaattcaat gactatacag gctatatctc aggcattcgg tggaaattat gaaacactgc 7380
taagaacatt gggttacgct acagaagact ttgatgatct tctagaaagt gacagcataa 7440
caggtcaaat catctatgtt gatctaagta gctactatat aattgtcagg gtttattttc 7500
ctattctgac tgaaattcaa caggcctata tccaagagtt gttaccagtg agcttcaaca 7560
atgataattc agaatggatc agtattgtcc caaatttcat attggtaagg aatacattaa 7620
tatcaaatat agagattgga ttttgcctaa ttacaaagag gagcgtgatc tgcaaccaag 7680
attatgccac acctatgacc aacaacatga gagaatgttt aacgggatcg actgagaagt 7740
gtcctcgaga gctggttgtt tcatcacatg ttcccagatt tgcactatct aacggggttc 7800
tgtttgccaa ttgcataagt gttacatgtc agtgtcaaac aacaggcagg gcaatctcac 7860
aatcaggaga acaaactctg ctgatgattg acaacaccac ctgtcctaca gccgtactcg 7920
gtaatgtgat tatcagctta gggaaatatc tggggtcagt aaattataat tctgaaggca 7980
ttgctatcgg tcctccagtc tttacagata aagttgatat atcaagtcag atatccagca 8040
tgaatcagtc cttacaacag tctaaggact atatcaaaga ggctcaacga ctccttgata 8100
ctgttaatcc atcattaata agcatgttgt ctatgatcat actgtatgta ttatcgatcg 8160
catcgttgtg tatagggttg attacattta tcagttttat cattgttgag aaaaagagaa 8220
acacctacag cagattagag gataggagag tcagacctac aagcagtggg gatctctact 8280
acattgggac atagtgtatt cagattgatg aaattatgtt agagaaatca gaaaacttct 8340
gactttcaga aatggattgt atacaattag ttagatcatc ctgaataatc gaggtgagaa 8400
cattgcaact ataaaatcag atcatgtaaa tagttgtaaa aaattaaaag cttcttttaa 8460
ttcttttgaa caataattta attaatatat aacatattct ctcacacgag cgctaaccta 8520
tacactctct actaatattt tatactcata attaatgata taatgacaaa taaggattca 8580
aattggatta tgatatagtt tcatactaca atagcatttc gaccaagaaa atatccttac 8640
aattatacaa tgtacttaac cgtgaatatg taattgataa tttcccttta gaaatttaat 8700
aaaaaactta ggacccaggt ccataactca ttggatactt aactgtatct ttctaagcta 8760
tcacatatca aaggagagat tgaatgcttt tttggagatc tagatcatta ctatatgtgt 8820
ctcctataat cacatcatag gagtgaacca taatacacat ctttgggtag gggaaggaaa 8880
gtattgttga cgtactgatt gatctgcttg agtcaaataa tcagtcataa caattcaaga 8940
aaatgccggc agaaaacaag aaagttagat tcgaaaatac tacttcagac aaagggaaaa 9000
ttcctagtaa agttattaag agctactacg gaaccatgga cattaagaaa ataaatgaag 9060
gattattgga cagcaaaata ttaagtgctt tcaacacagt aatagcattg cttggatcta 9120
tcgtgatcat agtgatgaat ataatgatca tccaaaatta cacaagatca acagacaatc 9180
aggccgtgat caaagatgcg ttgcagggta tccaacagca gatcaaaggg cttgctgaca 9240
aaatcggcac agagataggg cccaaagtat cactgattga cacatccagt accattacta 9300
tcccagctaa cattgggctg ttaggttcaa agatcagcca gtcgactgca agtataaatg 9360
agaatgtgaa tgaaaaatgc aaattcacac tacctccctt gaaaatccac gaatgtaaca 9420
tttcttgtcc taacccactc ccttttagag agtataggcc acagacagaa ggggtgagca 9480
atctagtagg attacctaat aatatttgcc tgcaaaagac atctaatcag atattgaagc 9540
caaagctgat ttcatacact ttacccgtag tcggtcaaag tggtacctgt atcacagacc 9600
cattgctggc tatggacgag ggctattttg catatagcca cctggaaaga atcggatcat 9660
gttcaagagg ggtctccaaa caaagaataa taggagttgg agaggtacta gacagaggtg 9720
atgaagttcc ttctttattt atgaccaatg tctggacccc accaaatcca aacaccgttt 9780
accactgtag tgctgtatac aacaatgaat tctattatgt actttgtgca gtgtcaactg 9840
ttggagaccc tattctgaat agcacctact ggtccggatc tctaatgatg acccgtctag 9900
ctgtgaaacc caagagtaat ggtgggggtt acaatcaaca tcaacttgcc ctacgaagta 9960
tcgagaaagg gaggtatgat aaagttatgc cgtatggacc ttcaggcatc aaacagggtg 10020
acaccctgta ttttcctgct gtaggatttt tggtcaggac agagtttaaa tacaatgatt 10080
caaattgtcc catcacgaag tgtcaataca gtaaacctga aaattgcagg ctatctatgg 10140
ggattagacc aaacagccat tatatccttc gatctggact attaaaatac aatctatcag 10200
atggggagaa ccccaaagtt gtattcattg aaatatctga tcaaagatta tctattggat 10260
ctcctagcaa aatctatgat tctttgggtc aacctgtttt ctaccaagcg tcattttcat 10320
gggatactat gattaaattt ggagatgttc taacagtcaa ccctctggtt gtcaattggc 10380
gtaataacac ggtaatatca agacccgggc aatcacaatg ccctagattc aatacatgtc 10440
cagagatctg ctgggaagga gtttataatg atgcattcct aattgacaga atcaattgga 10500
taagcgcggg tgtattcctt gacagcaatc agaccgcaga aaatcctgtt tttactgtat 10560
tcaaagataa tgaaatactt tatagggcac aactggcttc tgaggacacc aatgcacaaa 10620
aaacaataac taattgtttt ctcttgaaga ataagatttg gtgcatatca ttggttgaga 10680
tatatgacac aggagacaat gtcataagac ccaaactatt cgcggttaag ataccagagc 10740
aatgtacata aaaatcaacc tcataattta atggattgat ctaatataat gataataatc 10800
gtacaaagac atgtgatgta aacaaaattg ttgtaattaa ataagtcctc agctgaatac 10860
ttttttaaga ttagcaatag catgtttttc cagttattgg atagttgata atataattct 10920
gaaactgggt taataaataa tcttgatcgg tgatctttga gaacaatgat atcatatagt 10980
tcatcaagtg ataatcaatt ctttatatgt acactttaga gtatattttg agacttagta 11040
ttttcggccc gaatgttaaa tttaatagtt catacataac ctaaactcaa gttctaagca 11100
taatgataac aattaatgcg aacttgtctt gatgtaagga agatttgata ttaactgaga 11160
ctccacttga tatagtagag ctgaatcttg taaataaatt ataatgaata gtttattcaa 11220
agattatcat tcatattagt gtaaattaag aaaaacttag gacccaggtc cttgattatg 11280
ccaattttct cgagaaatca ttcaattgac catagactga aagcgttgtt acctagttct 11340
tcagaagaga tcttattaga attaatttat atgatctaat tcccttaaaa actgaatacc 11400
aaaaaacaaa aatggccgat gaattatcaa tatccgacat catttaccct gaatgtcatt 11460
tggatagtcc tatagtctct ggtaaactaa tatcagctat tgaatatgct caattgagac 11520
acaatcagcc cagtgatgat aaaagactgt ctgagaatat taggttaaac cttcacggga 11580
aaagaaagag tctatacata ttaagacaat ccaaacaggg tgattacatt agaaacaaca 11640
taaaaaacct aaaggaattc atgcatattg cgtaccctga atgcaataac attctattct 11700
ccatcacatc ccaaggcatg actagcaaac ttgataacat catgaaaaag tcattcaaag 11760
catacaatat cattagtaag aaagtaattg ggatgctgca aaatatcact agaaatctca 11820
taactcaaga tagaagagat gaaataatta atatacatga gtgtaggcga ttaggggatt 11880
tagggaagaa tatgagtcaa tctaaatggt atgagtgttt tttgttttgg tttactatca 11940
aaacagagat gcgagcagtg atcaagaatt cgcaaaagcc gaaattccgt tcagattcat 12000
gcataataca catgcgagac aaaagtactg aaataatcct aaatccgaat cttatctgca 12060
ttttcaaatc agacaaaact ggaaagaagt gttattatct tacacccgaa atggttctaa 12120
tgtattgtga tgtcctagag ggaaggatga tgatggagac aacagtcaaa tcggatatca 12180
agtaccaacc tctaatctcg agatccaatg ccctctgggg gctaattgat cccttgttcc 12240
ctgtcatggg aaacagaatt tacaatatag tgtctatgat agagccttta gttcttgcac 12300
tactccaact caaggatgag gctaggatcc tgaggggtgc atttctgcat cactgcataa 12360
aggaaatgca tcaagaattg agtgagtgtg gttttacaga tcagaagatt cggtctatgt 12420
ttattgatga tcttttatcc attctaaata tcgataatat acatctgttg gcagagttct 12480
tttctttctt tcgtacgttt ggccatccta ttcttgaggc taaagttgct gcagaaaaag 12540
tgagagaaca tatgttggca gataaagttc ttgaatatgc ccctataatg aaagcacatg 12600
ctatattctg cgggactata ataaatgggt atagggatag acacggagga gcctggcctc 12660
ctctttacct ccccgcacat gcatctaaac atataatccg tttgaaaaat tctggggaat 12720
ctttgaccat tgatgactgt gtcaagaatt gggaatcatt ctgtgggatt caatttgatt 12780
gtttcatgga gctgaaattg gacagtgatc tgagtatgta tatgaaagat aaagctttat 12840
ctccaatcaa agacgaatgg gacagtgtat acccacgtga agtgttgagc tataccccac 12900
cgaagtcaac cgagccaaga agattggttg acgtttttgt aaatgatgaa aactttgatc 12960
catacaacat gctggaatat gtcttatccg gtgcttatct cgaggatgaa caattcaatg 13020
tttcttatag cttgaaggag aaagagacga agcaagctgg acgattgttc gcaaagatga 13080
cctacaaaat gcgtgcatgt caagtcatag cagaggccct gatagcctca ggtgtcggta 13140
aatattttaa ggagaacggg atggttaagg atgagcacga acttttgaag acactcttcc 13200
aattgtctat ttcctcagtt cctcgaggga acagtcaggg taatgatcct caatccatca 13260
ataatataga aagagatttc caatacttta aaggggtcac taccaatgtg aaagacaaaa 13320
agaataactc ttttaataag gttaaatctg ctctcaataa tccgtgccaa gctgacggag 13380
tccatcataa catgtcaccc aatacacgaa atcgttataa gtgtagtaat acaagtaagt 13440
cttttctcga ttatcatacc gagtttaatc ctcacaatca ctataaatca gacaatacag 13500
aggcggccgt actgtccagg tatgaggaca acactgggac aaaatttgat acagtaagtg 13560
catttcttac aactgatctt aagaaattct gtctcaattg gagatacgaa tcaatggcta 13620
tatttgctga acgtctggat gagatatacg gtttacctgg attttttaat tggatgcaca 13680
aacgactaga aagatctgtt atctatgttg cagaccctaa ttgcccccct aatattgaca 13740
aacatatgga actagaaaaa actcctgaag atgatatatt cattcattat cctaaaggcg 13800
gtattgaagg atatagccaa aaaacatgga ctatagcaac tatccccttt ttattcttga 13860
gtgcctatga gacaaacacg aggattgctg caattgtcca aggagacaat gaatcaattg 13920
ctatcactca aaaagttcat cctaatcttc cctacaaggt aaagaaagag atctgtgcaa 13980
agcaagctca gctttatttt gaaaggttaa ggatgaactt aagagccctc ggccacaatc 14040
ttaaagctac agaaactatc atcagtacac atctttttat ttattcgaag aaaattcatt 14100
atgatggtgc tgtgctgtct caggcactca aatcaatgtc aagatgttgc ttttggtcag 14160
agactctggt ggatgaaact agatcagctt gtagtaacat cagcactaca atagctaaag 14220
ctatagaaaa tgggttgtca agaaatgtcg gctattgcat caatattttg aaagtaattc 14280
agcagcttct catatcaact gagtttagta ttaacgagac attgacactg gatgtgacat 14340
ctcccatttc aaataattta gattggctta taacagctgc attaatcccg gcacctattg 14400
gaggattcaa ttaccttaat ttgtctagaa tttttgttag aaatataggt gatccggtta 14460
cagcatcttt ggctgatctt aagagaatga ttgatcacag tattatgact gaaagcgtat 14520
tacaaaaagt tatgaatcaa gaacctggtg atgcgagttt cttggactgg gccagtgatc 14580
catactcggg caacttgcct gactcacaaa gcatcactaa aacaattaaa aatatcacag 14640
caaggactat actgaggaac tcaccgaacc caatgctaaa aggtttattt catgacaaat 14700
cttttgatga agatcttgaa ctagctagct tcttaatgga caggagggtt atattaccta 14760
gagccgctca tgagatactg gataattcat tgacaggtgc cagagaggaa attgctggtt 14820
tattagatac aactaaaggc ttgatcagat cagggctaag aaagagtgga cttcagccaa 14880
agttagtttc tagattatct catcatgatt ataatcaatt tttaatactg aacaaacttc 14940
tatcaaacag aagacaaaat gacttgatat catcaaatac ttgctcagtt gacttggcac 15000
gagcattgag atctcacatg tggagggaat tagcgttagg tagagtaata tacggtcttg 15060
aggtaccaga tgcacttgag gctatggtgg gaaggtatat aacagggagc ttagagtgcc 15120
aaatttgtga gcagggaaac acgatgtatg ggtggttctt tgtacctagg gattcccaat 15180
tggatcaggt agatagagag cactcatcaa taagagtacc ttatgtagga tcaagtacgg 15240
atgaaagatc ggatatcaaa ctagggaatg tcaaaagacc aactaaggcc ttgcgttctg 15300
ctatcagaat tgcgacagta tatacttggg cctatgggga caatgaagag tgttggtatg 15360
aagcttggta cctagcgtct cagagggtaa acatagactt agatgtattg aaagctataa 15420
ccccagtttc cacttcaaac aatttatccc atagattgag agataaatcc acacaattta 15480
agtttgcagg gagtgtactc aacagagttt ctagatatgt taacataagc aatgacaatc 15540
tagatttcag aattgaggga gaaaaggtag atacgaatct tatttatcaa caagcaatgc 15600
tattagggtt atcggtattg gaaggtaaat tcagattgag attagaaact gatgattaca 15660
acgggatata tcacttacac gtaaaggata attgttgtgt caaagaagtg gctgatgtag 15720
gccaagtaga cgctgagttg cctatcccag aatatactga agtggataac aatcatctta 15780
tatatgatcc agaccccgtt tcagaaatag attgcagccg tctttctaat caggagtcca 15840
aatcaagaga attagacttt cctttatggt caactgagga acttcatgat gtcctagcta 15900
agactgttgc tcagaccgtt cttgagatta taacaaaggc tgacaaggat gttttaaagc 15960
aacaccttgc aatagactct gacgataaca tcaacagctt aatcacagaa tttctaatag 16020
ttgatcctga actgtttgca ctttatctag gacaatctat atcaataaaa tgggcctttg 16080
aaattcatca taggcgtcct agaggaagac atactatggt cgacttattg tcagatcttg 16140
tatcaaatac atcaaagcac acttacaaag tgttgtcaaa tgccttgtca catcctagag 16200
tattcaagag atttgtaaac tgtggcttgc tattgcctac acagggtcct taccttcatc 16260
aacaagattt tgaaaagttg tctcaaaacc ttcttgtaac atcttatatg atttatctaa 16320
tgaactggtg tgacttcaag aaattcccct ttttaatcgc cgaacaggat gaaactgtga 16380
taagtctacg agaggatata ataacatcca aacatctctg tgttataatt gacttatatg 16440
caaatcacca taaacctcct tggataatag atctaaaccc acaagaaaaa atatgtgtac 16500
tgcgtgactt tatttctaaa tctaggcatg tggacacgtc ctccagatca tggaatactt 16560
ctgacctgga ttttgtaata ttctatgcat ctttgactta tttgagaaga ggtataataa 16620
aacaattaag gataagacaa gttactgagg ttatagatac cacaacaatg ttaagggaca 16680
atataattgt agagaatcct cctattaaaa caggagtgtt agacatcaga ggttgtataa 16740
tatacaattt agaggaaatc ctgtctatga acacaaaatc agcatcaaaa aagatcttta 16800
atcttaatag taggccgtca gtggagaatc ataaatatag aaggataggt ctcaactcat 16860
catcttgtta caaggcatta aatctatcac ctctgattca aaggtatttg ccgtcgggag 16920
ctcaaaggtt gtttatagga gaaggttctg ggagcatgat gttattatat cagtctacat 16980
tggggcaatc aatttctttt tacaattcag gtatagatgg agattatata ccaggtcaaa 17040
gagaactgaa actatttccc tctgaatact caattgctga ggaagaccca tctctgacgg 17100
ggaaattgaa aggactagtg gtgcccctat tcaatggaag accagaaaca acatggatcg 17160
ggaatttaga ctcctacgag tatatcataa ataggacagc ggggcgaagt ataggtcttg 17220
tccattctga catggagtct gggattgaca aaaatgtaga ggagatacta gtagaacatt 17280
cccatctaat atctatcgcg ataaatgtta tgatggagga cggactatta gtatccaaga 17340
tagcatacac ccctggattc ccaatctcaa gattatttaa catgtacaga tcatatttcg 17400
gactagtact ggtgtgtttc ccagtatata gtaatccaga ttctactgaa gtatatcttc 17460
tttgcttaca gaagacggtc aagactattg ttcccccgca aaaagtcctt gagcactcta 17520
atttgcacga tgaagtcaat gaccagggaa taacatcagt gatttttaaa atcaagaatt 17580
cacagtctaa gcagttccac gatgatctaa agaagtacta tcagattgac caaccttttt 17640
ttgtaccaac taaaatcact agtgatgaac aagtacttct ccaagcaggg ctgaaactca 17700
atgggccaga aattcttaag agtgaaatca gttatgatat cggttcagat atcaatacat 17760
taagagacac catcataatt atgttaaatg aggctatgaa ttattttgat gacaacagat 17820
caccttcaca ccacctagaa ccctatccag ttttggagag aactagaatt aaaacaataa 17880
tgaattgtgt gactaaaaaa gtgattgtct actcacttat caagttcaag gacaccaaaa 17940
gctcagaact ttatcacatc aaaaataaca tcagaagaaa agttctaatc ttagatttca 18000
gatcgaagct catgacaaag actctaccta aagggatgca agagagaaga gaaaaaaacg 18060
gtttcaaaga agtttggata gtagatttat cgaatcgaga agttaaaatc tggtggaaga 18120
taatcggata catatctatt atctgattta accttccaaa tccaagacca actgataact 18180
tatgttgatc taaggttcag ttattaagaa aaacttaata acgattcttc tttacccttg 18240
ttcggt 18246
<210>18
<211>2244
<212>PRT
<213>Nipah virus
<400>18
Met Ala Asp Glu Leu Ser Ile Ser Asp Ile Ile Tyr Pro Glu Cys His
1 5 10 15
Leu Asp Ser Pro Ile Val Ser Gly Lys Leu Ile Ser Ala Ile Glu Tyr
20 25 30
Ala Gln Leu Arg His Asn Gln Pro Ser Asp Asp Lys Arg Leu Ser Glu
35 40 45
Asn Ile Arg Leu Asn Leu His Gly Lys Arg Lys Ser Leu Tyr Ile Leu
50 55 60
Arg Gln Ser Lys Gln Gly Asp Tyr Ile Arg Asn Asn Ile Lys Asn Leu
65 70 75 80
Lys Glu Phe Met His Ile Ala Tyr Pro Glu Cys Asn Asn Ile Leu Phe
85 90 95
Ser Ile Thr Ser Gln Gly Met Thr Ser Lys Leu Asp Asn Ile Met Lys
100 105 110
Lys Ser Phe Lys Ala Tyr Asn Ile Ile Ser Lys Lys Val Ile Gly Met
115 120 125
Leu Gln Asn Ile Thr Arg Asn Leu Ile Thr Gln Asp Arg Arg Asp Glu
130 135 140
Ile Ile Asn Ile His Glu Cys Arg Arg Leu Gly Asp Leu Gly Lys Asn
145 150 155 160
Met Ser Gln Ser Lys Trp Tyr Glu Cys Phe Leu Phe Trp Phe Thr Ile
165 170 175
Lys Thr Glu Met Arg Ala Val Ile Lys Asn Ser Gln Lys Pro Lys Phe
180 185 190
Arg Ser Asp Ser Cys Ile Ile His Met Arg Asp Lys Ser Thr Glu Ile
195 200 205
Ile Leu Asn Pro Asn Leu Ile Cys Ile Phe Lys Ser Asp Lys Thr Gly
210 215 220
Lys Lys Cys Tyr Tyr Leu Thr Pro Glu Met Val Leu Met Tyr Cys Asp
225 230 235 240
Val Leu Glu Gly Arg Met Met Met Glu Thr Thr Val Lys Ser Asp Ile
245 250 255
Lys Tyr Gln Pro Leu Ile Ser Arg Ser Asn Ala Leu Trp Gly Leu Ile
260 265 270
Asp Pro Leu Phe Pro Val Met Gly Asn Arg Ile Tyr Asn Ile Val Ser
275 280 285
Met Ile Glu Pro Leu Val Leu Ala Leu Leu Gln Leu Lys Asp Glu Ala
290 295 300
Arg Ile Leu Arg Gly Ala Phe Leu His His Cys Ile Lys Glu Met His
305 310 315 320
Gln Glu Leu Ser Glu Cys Gly Phe Thr Asp Gln Lys Ile Arg Ser Met
325 330 335
Phe Ile Asp Asp Leu Leu Ser Ile Leu Asn Ile Asp Asn Ile His Leu
340 345 350
Leu Ala Glu Phe Phe Ser Phe Phe Arg Thr Phe Gly His Pro Ile Leu
355 360 365
Glu Ala Lys Val Ala Ala Glu Lys Val Arg Glu His Met Leu Ala Asp
370 375 380
Lys Val Leu Glu Tyr Ala Pro Ile Met Lys Ala His Ala Ile Phe Cys
385 390 395 400
Gly Thr Ile Ile Asn Gly Tyr Arg Asp Arg His Gly Gly Ala Trp Pro
405 410 415
Pro Leu Tyr Leu Pro Ala His Ala Ser Lys His Ile Ile Arg Leu Lys
420 425 430
Asn Ser Gly Glu Ser Leu Thr Ile Asp Asp Cys Val Lys Asn Trp Glu
435 440 445
Ser Phe Cys Gly Ile Gln Phe Asp Cys Phe Met Glu Leu Lys Leu Asp
450 455 460
Ser Asp Leu Ser Met Tyr Met Lys Asp Lys Ala Leu Ser Pro Ile Lys
465 470 475 480
Asp Glu Trp Asp Ser Val Tyr Pro Arg Glu Val Leu Ser Tyr Thr Pro
485 490 495
Pro Lys Ser Thr Glu Pro Arg Arg Leu Val Asp Val Phe Val Asn Asp
500 505 510
Glu Asn Phe Asp Pro Tyr Asn Met Leu Glu Tyr Val Leu Ser Gly Ala
515 520 525
Tyr Leu Glu Asp Glu Gln Phe Asn Val Ser Tyr Ser Leu Lys Glu Lys
530 535 540
Glu Thr Lys Gln Ala Gly Arg Leu Phe Ala Lys Met Thr Tyr Lys Met
545 550 555 560
Arg Ala Cys Gln Val Ile Ala Glu Ala Leu Ile Ala Ser Gly Val Gly
565 570 575
Lys Tyr Phe Lys Glu Asn Gly Met Val Lys Asp Glu His Glu Leu Leu
580 585 590
Lys Thr Leu Phe Gln Leu Ser Ile Ser Ser Val Pro Arg Gly Asn Ser
595 600 605
Gln Gly Asn Asp Pro Gln Ser Ile Asn Asn Ile Glu Arg Asp Phe Gln
610 615 620
Tyr Phe Lys Gly Val Thr Thr Asn Val Lys Asp Lys Lys Asn Asn Ser
625 630 635 640
Phe Asn Lys Val Lys Ser Ala Leu Asn Asn Pro Cys Gln Ala Asp Gly
645 650 655
Val His His Asn Met Ser Pro Asn Thr Arg Asn Arg Tyr Lys Cys Ser
660 665 670
Asn Thr Ser Lys Ser Phe Leu Asp Tyr His Thr Glu Phe Asn Pro His
675 680 685
Asn His Tyr Lys Ser Asp Asn Thr Glu Ala Ala Val Leu Ser Arg Tyr
690 695 700
Glu Asp Asn Thr Gly Thr Lys Phe Asp Thr Val Ser Ala Phe Leu Thr
705 710 715 720
Thr Asp Leu Lys Lys Phe Cys Leu Asn Trp Arg Tyr Glu Ser Met Ala
725 730 735
Ile Phe Ala Glu Arg Leu Asp Glu Ile Tyr Gly Leu Pro Gly Phe Phe
740 745 750
Asn Trp Met His Lys Arg Leu Glu Arg Ser Val Ile Tyr Val Ala Asp
755 760 765
Pro Asn Cys Pro Pro Asn Ile Asp Lys His Met Glu Leu Glu Lys Thr
770 775 780
Pro Glu Asp Asp Ile Phe Ile His Tyr Pro Lys Gly Gly Ile Glu Gly
785 790 795 800
Tyr Ser Gln Lys Thr Trp Thr Ile Ala Thr Ile Pro Phe Leu Phe Leu
805 810 815
Ser Ala Tyr Glu Thr Asn Thr Arg Ile Ala Ala Ile Val Gln Gly Asp
820 825 830
Asn Glu Ser Ile Ala Ile Thr Gln Lys Val His Pro Asn Leu Pro Tyr
835 840 845
Lys Val Lys Lys Glu Ile Cys Ala Lys Gln Ala Gln Leu Tyr Phe Glu
850 855 860
Arg Leu Arg Met Asn Leu Arg Ala Leu Gly His Asn Leu Lys Ala Thr
865 870 875 880
Glu Thr Ile Ile Ser Thr His Leu Phe Ile Tyr Ser Lys Lys Ile His
885 890 895
Tyr Asp Gly Ala Val Leu Ser Gln Ala Leu Lys Ser Met Ser Arg Cys
900 905 910
Cys Phe Trp Ser Glu Thr Leu Val Asp Glu Thr Arg Ser Ala Cys Ser
915 920 925
Asn Ile Ser Thr Thr Ile Ala Lys Ala Ile Glu Asn Gly Leu Ser Arg
930 935 940
Asn Val Gly Tyr Cys Ile Asn Ile Leu Lys Val Ile Gln Gln Leu Leu
945 950 955 960
Ile Ser Thr Glu Phe Ser Ile Asn Glu Thr Leu Thr Leu Asp Val Thr
965 970 975
Ser Pro Ile Ser Asn Asn Leu Asp Trp Leu Ile Thr Ala Ala Leu Ile
980 985 990
Pro Ala Pro Ile Gly Gly Phe Asn Tyr Leu Asn Leu Ser Arg Ile Phe
995 1000 1005
Val Arg Asn Ile Gly Asp Pro Val Thr Ala Ser Leu Ala Asp Leu
1010 1015 1020
Lys Arg Met Ile Asp His Ser Ile Met Thr Glu Ser Val Leu Gln
1025 1030 1035
Lys Val Met Asn Gln Glu Pro Gly Asp Ala Ser Phe Leu Asp Trp
1040 1045 1050
Ala Ser Asp Pro Tyr Ser Gly Asn Leu Pro Asp Ser Gln Ser Ile
1055 1060 1065
Thr Lys Thr Ile Lys Asn Ile Thr Ala Arg Thr Ile Leu Arg Asn
1070 1075 1080
Ser Pro Asn Pro Met Leu Lys Gly Leu Phe His Asp Lys Ser Phe
1085 1090 1095
Asp Glu Asp Leu Glu Leu Ala Ser Phe Leu Met Asp Arg Arg Val
1100 1105 1110
Ile Leu Pro Arg Ala Ala His Glu Ile Leu Asp Asn Ser Leu Thr
1115 1120 1125
Gly Ala Arg Glu Glu Ile Ala Gly Leu Leu Asp Thr Thr Lys Gly
1130 1135 1140
Leu Ile Arg Ser Gly Leu Arg Lys Ser Gly Leu Gln Pro Lys Leu
1145 1150 1155
Val Ser Arg Leu Ser His His Asp Tyr Asn Gln Phe Leu Ile Leu
1160 1165 1170
Asn Lys Leu Leu Ser Asn Arg Arg Gln Asn Asp Leu Ile Ser Ser
1175 1180 1185
Asn Thr Cys Ser Val Asp Leu Ala Arg Ala Leu Arg Ser His Met
1190 1195 1200
Trp Arg Glu Leu Ala Leu Gly Arg Val Ile Tyr Gly Leu Glu Val
1205 1210 1215
Pro Asp Ala Leu Glu Ala Met Val Gly Arg Tyr Ile Thr Gly Ser
1220 1225 1230
Leu Glu Cys Gln Ile Cys Glu Gln Gly Asn Thr Met Tyr Gly Trp
1235 1240 1245
Phe Phe Val Pro Arg Asp Ser Gln Leu Asp Gln Val Asp Arg Glu
1250 1255 1260
His Ser Ser Ile Arg Val Pro Tyr Val Gly Ser Ser Thr Asp Glu
1265 1270 1275
Arg Ser Asp Ile Lys Leu Gly Asn Val Lys Arg Pro Thr Lys Ala
1280 1285 1290
Leu Arg Ser Ala Ile Arg Ile Ala Thr Val Tyr Thr Trp Ala Tyr
1295 1300 1305
Gly Asp Asn Glu Glu Cys Trp Tyr Glu Ala Trp Tyr Leu Ala Ser
1310 1315 1320
Gln Arg Val Asn Ile Asp Leu Asp Val Leu Lys Ala Ile Thr Pro
1325 1330 1335
Val Ser Thr Ser Asn Asn Leu Ser His Arg Leu Arg Asp Lys Ser
1340 1345 1350
Thr Gln Phe Lys Phe Ala Gly Ser Val Leu Asn Arg Val Ser Arg
1355 1360 1365
Tyr Val Asn Ile Ser Asn Asp Asn Leu Asp Phe Arg Ile Glu Gly
1370 1375 1380
Glu Lys Val Asp Thr Asn Leu Ile Tyr Gln Gln Ala Met Leu Leu
1385 1390 1395
Gly Leu Ser Val Leu Glu Gly Lys Phe Arg Leu Arg Leu Glu Thr
1400 1405 1410
Asp Asp Tyr Asn Gly Ile Tyr His Leu His Val Lys Asp Asn Cys
1415 1420 1425
Cys Val Lys Glu Val Ala Asp Val Gly Gln Val Asp Ala Glu Leu
1430 1435 1440
Pro Ile Pro Glu Tyr Thr Glu Val Asp Asn Asn His Leu Ile Tyr
1445 1450 1455
Asp Pro Asp Pro Val Ser Glu Ile Asp Cys Ser Arg Leu Ser Asn
1460 1465 1470
Gln Glu Ser Lys Ser Arg Glu Leu Asp Phe Pro Leu Trp Ser Thr
1475 1480 1485
Glu Glu Leu His Asp Val Leu Ala Lys Thr Val Ala Gln Thr Val
1490 1495 1500
Leu Glu Ile Ile Thr Lys Ala Asp Lys Asp Val Leu Lys Gln His
1505 1510 1515
Leu Ala Ile Asp Ser Asp Asp Asn Ile Asn Ser Leu Ile Thr Glu
1520 1525 1530
Phe Leu Ile Val Asp Pro Glu Leu Phe Ala Leu Tyr Leu Gly Gln
1535 1540 1545
Ser Ile Ser Ile Lys Trp Ala Phe Glu Ile His His Arg Arg Pro
1550 1555 1560
Arg Gly Arg His Thr Met Val Asp Leu Leu Ser Asp Leu Val Ser
1565 1570 1575
Asn Thr Ser Lys His Thr Tyr Lys Val Leu Ser Asn Ala Leu Ser
1580 1585 1590
His Pro Arg Val Phe Lys Arg Phe Val Asn Cys Gly Leu Leu Leu
1595 1600 1605
Pro Thr Gln Gly Pro Tyr Leu His Gln Gln Asp Phe Glu Lys Leu
1610 1615 1620
Ser Gln Asn Leu Leu Val Thr Ser Tyr Met Ile Tyr Leu Met Asn
1625 1630 1635
Trp Cys Asp Phe Lys Lys Phe Pro Phe Leu Ile Ala Glu Gln Asp
1640 1645 1650
Glu Thr Val Ile Ser Leu Arg Glu Asp Ile Ile Thr Ser Lys His
1655 1660 1665
Leu Cys Val Ile Ile Asp Leu Tyr Ala Asn His His Lys Pro Pro
1670 1675 1680
Trp Ile Ile Asp Leu Asn Pro Gln Glu Lys Ile Cys Val Leu Arg
1685 1690 1695
Asp Phe Ile Ser Lys Ser Arg His Val Asp Thr Ser Ser Arg Ser
1700 1705 1710
Trp Asn Thr Ser Asp Leu Asp Phe Val Ile Phe Tyr Ala Ser Leu
1715 1720 1725
Thr Tyr Leu Arg Arg Gly Ile Ile Lys Gln Leu Arg Ile Arg Gln
1730 1735 1740
Val Thr Glu Val Ile Asp Thr Thr Thr Met Leu Arg Asp Asn Ile
1745 1750 1755
Ile Val Glu Asn Pro Pro Ile Lys Thr Gly Val Leu Asp Ile Arg
1760 1765 1770
Gly Cys Ile Ile Tyr Asn Leu Glu Glu Ile Leu Ser Met Asn Thr
1775 1780 1785
Lys Ser Ala Ser Lys Lys Ile Phe Asn Leu Asn Ser Arg Pro Ser
1790 1795 1800
Val Glu Asn His Lys Tyr Arg Arg Ile Gly Leu Asn Ser Ser Ser
1805 1810 1815
Cys Tyr Lys Ala Leu Asn Leu Ser Pro Leu Ile Gln Arg Tyr Leu
1820 1825 1830
Pro Ser Gly Ala Gln Arg Leu Phe Ile Gly Glu Gly Ser Gly Ser
1835 1840 1845
Met Met Leu Leu Tyr Gln Ser Thr Leu Gly Gln Ser Ile Ser Phe
1850 1855 1860
Tyr Asn Ser Gly Ile Asp Gly Asp Tyr Ile Pro Gly Gln Arg Glu
1865 1870 1875
Leu Lys Leu Phe Pro Ser Glu Tyr Ser Ile Ala Glu Glu Asp Pro
1880 1885 1890
Ser Leu Thr Gly Lys Leu Lys Gly Leu Val Val Pro Leu Phe Asn
1895 1900 1905
Gly Arg Pro Glu Thr Thr Trp Ile Gly Asn Leu Asp Ser Tyr Glu
1910 1915 1920
Tyr Ile Ile Asn Arg Thr Ala Gly Arg Ser Ile Gly Leu Val His
1925 1930 1935
Ser Asp Met Glu Ser Gly Ile Asp Lys Asn Val Glu Glu Ile Leu
1940 1945 1950
Val Glu His Ser His Leu Ile Ser Ile Ala Ile Asn Val Met Met
1955 1960 1965
Glu Asp Gly Leu Leu Val Ser Lys Ile Ala Tyr Thr Pro Gly Phe
1970 1975 1980
Pro Ile Ser Arg Leu Phe Asn Met Tyr Arg Ser Tyr Phe Gly Leu
1985 1990 1995
Val Leu Val Cys Phe Pro Val Tyr Ser Asn Pro Asp Ser Thr Glu
2000 2005 2010
Val Tyr Leu Leu Cys Leu Gln Lys Thr Val Lys Thr Ile Val Pro
2015 2020 2025
Pro Gln Lys Val Leu Glu His Ser Asn Leu His Asp Glu Val Asn
2030 2035 2040
Asp Gln Gly Ile Thr Ser Val Ile Phe Lys Ile Lys Asn Ser Gln
2045 2050 2055
Ser Lys Gln Phe His Asp Asp Leu Lys Lys Tyr Tyr Gln Ile Asp
2060 2065 2070
Gln Pro Phe Phe Val Pro Thr Lys Ile Thr Ser Asp Glu Gln Val
2075 2080 2085
Leu Leu Gln Ala Gly Leu Lys Leu Asn Gly Pro Glu Ile Leu Lys
2090 2095 2100
Ser Glu Ile Ser Tyr Asp Ile Gly Ser Asp Ile Asn Thr Leu Arg
2105 2110 2115
Asp Thr Ile Ile Ile Met Leu Asn Glu Ala Met Asn Tyr Phe Asp
2120 2125 2130
Asp Asn Arg Ser Pro Ser His His Leu Glu Pro Tyr Pro Val Leu
2135 2140 2145
Glu Arg Thr Arg Ile Lys Thr Ile Met Asn Cys Val Thr Lys Lys
2150 2155 2160
Val Ile Val Tyr Ser Leu Ile Lys Phe Lys Asp Thr Lys Ser Ser
2165 2170 2175
Glu Leu Tyr His Ile Lys Asn Asn Ile Arg Arg Lys Val Leu Ile
2180 2185 2190
Leu Asp Phe Arg Ser Lys Leu Met Thr Lys Thr Leu Pro Lys Gly
2195 2200 2205
Met Gln Glu Arg Arg Glu Lys Asn Gly Phe Lys Glu Val Trp Ile
2210 2215 2220
Val Asp Leu Ser Asn Arg Glu Val Lys Ile Trp Trp Lys Ile Ile
2225 2230 2235
Gly Tyr Ile Ser Ile Ile
2240
<210>19
<211>602
<212>PRT
<213>Nipah virus
<400>19
Met Pro Ala Glu Asn Lys Lys Val Arg Phe Glu Asn Thr Thr Ser Asp
1 5 10 15
Lys Gly Lys Ile Pro Ser Lys Val Ile Lys Ser Tyr Tyr Gly Thr Met
20 25 30
Asp Ile Lys Lys Ile Asn Glu Gly Leu Leu Asp Ser Lys Ile Leu Ser
35 40 45
Ala Phe Asn Thr Val Ile Ala Leu Leu Gly Ser Ile Val Ile Ile Val
50 55 60
Met Asn Ile Met Ile Ile Gln Asn Tyr Thr Arg Ser Thr Asp Asn Gln
65 70 75 80
Ala Val Ile Lys Asp Ala Leu Gln Gly Ile Gln Gln Gln Ile Lys Gly
85 90 95
Leu Ala Asp Lys Ile Gly Thr Glu Ile Gly Pro Lys Val Ser Leu Ile
100 105 110
Asp Thr Ser Ser Thr Ile Thr Ile Pro Ala Asn Ile Gly Leu Leu Gly
115 120 125
Ser Lys Ile Ser Gln Ser Thr Ala Ser Ile Asn Glu Asn Val Asn Glu
130 135 140
Lys Cys Lys Phe Thr Leu Pro Pro Leu Lys Ile His Glu Cys Asn Ile
145 150 155 160
Ser Cys Pro Asn Pro Leu Pro Phe Arg Glu Tyr Arg Pro Gln Thr Glu
165 170 175
Gly Val Ser Asn Leu Val Gly Leu Pro Asn Asn Ile Cys Leu Gln Lys
180 185 190
Thr Ser Asn Gln Ile Leu Lys Pro Lys Leu Ile Ser Tyr Thr Leu Pro
195 200 205
Val Val Gly Gln Ser Gly Thr Cys Ile Thr Asp Pro Leu Leu Ala Met
210 215 220
Asp Glu Gly Tyr Phe Ala Tyr Ser His Leu Glu Arg Ile Gly Ser Cys
225 230 235 240
Ser Arg Gly Val Ser Lys Gln Arg Ile Ile Gly Val Gly Glu Val Leu
245 250 255
Asp Arg Gly Asp Glu Val Pro Ser Leu Phe Met Thr Asn Val Trp Thr
260 265 270
Pro Pro Asn Pro Asn Thr Val Tyr His Cys Ser Ala Val Tyr Asn Asn
275 280 285
Glu Phe Tyr Tyr Val Leu Cys Ala Val Ser Thr Val Gly Asp Pro Ile
290 295 300
Leu Asn Ser Thr Tyr Trp Ser Gly Ser Leu Met Met Thr Arg Leu Ala
305 310 315 320
Val Lys Pro Lys Ser Asn Gly Gly Gly Tyr Asn Gln His Gln Leu Ala
325 330 335
Leu Arg Ser Ile Glu Lys Gly Arg Tyr Asp Lys Val Met Pro Tyr Gly
340 345 350
Pro Ser Gly Ile Lys Gln Gly Asp Thr Leu Tyr Phe Pro Ala Val Gly
355 360 365
Phe Leu Val Arg Thr Glu Phe Lys Tyr Asn Asp Ser Asn Cys Pro Ile
370 375 380
Thr Lys Cys Gln Tyr Ser Lys Pro Glu Asn Cys Arg Leu Ser Met Gly
385 390 395 400
Ile Arg Pro Asn Ser His Tyr Ile Leu Arg Ser Gly Leu Leu Lys Tyr
405 410 415
Asn Leu Ser Asp Gly Glu Asn Pro Lys Val Val Phe Ile Glu Ile Ser
420 425 430
Asp Gln Arg Leu Ser Ile Gly Ser Pro Ser Lys Ile Tyr Asp Ser Leu
435 440 445
Gly Gln Pro Val Phe Tyr Gln Ala Ser Phe Ser Trp Asp Thr Met Ile
450 455 460
Lys Phe Gly Asp Val Leu Thr Val Asn Pro Leu Val Val Asn Trp Arg
465 470 475 480
Asn Asn Thr Val Ile Ser Arg Pro Gly Gln Ser Gln Cys Pro Arg Phe
485 490 495
Asn Thr Cys Pro Glu Ile Cys Trp Glu Gly Val Tyr Asn Asp Ala Phe
500 505 510
Leu Ile Asp Arg Ile Asn Trp Ile Ser Ala Gly Val Phe Leu Asp Ser
515 520 525
Asn Gln Thr Ala Glu Asn Pro Val Phe Thr Val Phe Lys Asp Asn Glu
530 535 540
Ile Leu Tyr Arg Ala Gln Leu Ala Ser Glu Asp Thr Asn Ala Gln Lys
545 550 555 560
Thr Ile Thr Asn Cys Phe Leu Leu Lys Asn Lys Ile Trp Cys Ile Ser
565 570 575
Leu Val Glu Ile Tyr Asp Thr Gly Asp Asn Val Ile Arg Pro Lys Leu
580 585 590
Phe Ala Val Lys Ile Pro Glu Gln Cys Thr
595 600
<210>20
<211>546
<212>PRT
<213>Nipah virus
<400>20
Met Val Val Ile Leu Asp Lys Arg Cys Tyr Cys Asn Leu Leu Ile Leu
1 5 10 15
Ile Leu Met Ile Ser Glu Cys Ser Val Gly Ile Leu His Tyr Glu Lys
20 25 30
Leu Ser Lys Ile Gly Leu Val Lys Gly Val Thr Arg Lys Tyr Lys Ile
35 40 45
Lys Ser Asn Pro Leu Thr Lys Asp Ile Val Ile Lys Met Ile Pro Asn
50 55 60
Val Ser Asn Met Ser Gln Cys Thr Gly Ser Val Met Glu Asn Tyr Lys
65 70 75 80
Thr Arg Leu Asn Gly Ile Leu Thr Pro Ile Lys Gly Ala Leu Glu Ile
85 90 95
Tyr Lys Asn Asn Thr His Asp Leu Val Gly Asp Val Arg Leu Ala Gly
100 105 110
Val Ile Met Ala Gly Val Ala Ile Gly Ile Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Val Ala Leu Tyr Glu Ala Met Lys Asn Ala Asp Asn Ile
130 135 140
Asn Lys Leu Lys Ser Ser Ile Glu Ser Thr Asn Glu Ala Val Val Lys
145 150 155 160
Leu Gln Glu Thr Ala Glu Lys Thr Val Tyr Val Leu Thr Ala Leu Gln
165 170 175
Asp Tyr Ile Asn Thr Asn Leu Val Pro Thr Ile Asp Lys Ile Ser Cys
180 185 190
Lys Gln Thr Glu Leu Ser Leu Asp Leu Ala Leu Ser Lys Tyr Leu Ser
195 200 205
Asp Leu Leu Phe Val Phe Gly Pro Asn Leu Gln Asp Pro Val Ser Asn
210 215 220
Ser Met Thr Ile Gln Ala Ile Ser Gln Ala Phe Gly Gly Asn Tyr Glu
225 230 235 240
Thr Leu Leu Arg Thr Leu Gly Tyr Ala Thr Glu Asp Phe Asp Asp Leu
245 250 255
Leu Glu Ser Asp Ser Ile Thr Gly Gln Ile Ile Tyr Val Asp Leu Ser
260 265 270
Ser Tyr Tyr Ile Ile Val Arg Val Tyr Phe Pro Ile Leu Thr Glu Ile
275 280 285
Gln Gln Ala Tyr Ile Gln Glu Leu Leu Pro Val Ser Phe Asn Asn Asp
290 295 300
Asn Ser Glu Trp Ile Ser Ile Val Pro Asn Phe Ile Leu Val Arg Asn
305 310 315 320
Thr Leu Ile Ser Asn Ile Glu Ile Gly Phe Cys Leu Ile Thr Lys Arg
325 330 335
Ser Val Ile Cys Asn Gln Asp Tyr Ala Thr Pro Met Thr Asn Asn Met
340 345 350
Arg Glu Cys Leu Thr Gly Ser Thr Glu Lys Cys Pro Arg Glu Leu Val
355 360 365
Val Ser Ser His Val Pro Arg Phe Ala Leu Ser Asn Gly Val Leu Phe
370 375 380
Ala Asn Cys Ile Ser Val Thr Cys Gln Cys Gln Thr Thr Gly Arg Ala
385 390 395 400
Ile Ser Gln Ser Gly Glu Gln Thr Leu Leu Met Ile Asp Asn Thr Thr
405 410 415
Cys Pro Thr Ala Val Leu Gly Asn Val Ile Ile Ser Leu Gly Lys Tyr
420 425 430
Leu Gly Ser Val Asn Tyr Asn Ser Glu Gly Ile Ala Ile Gly Pro Pro
435 440 445
Val Phe Thr Asp Lys Val Asp Ile Ser Ser Gln Ile Ser Ser Met Asn
450 455 460
Gln Ser Leu Gln Gln Ser Lys Asp Tyr Ile Lys Glu Ala Gln Arg Leu
465 470 475 480
Leu Asp Thr Val Asn Pro Ser Leu Ile Ser Met Leu Ser Met Ile Ile
485 490 495
Leu Tyr Val Leu Ser Ile Ala Ser Leu Cys Ile Gly Leu Ile Thr Phe
500 505 510
Ile Ser Phe Ile Ile Val Glu Lys Lys Arg Asn Thr Tyr Ser Arg Leu
515 520 525
Glu Asp Arg Arg Val Arg Pro Thr Ser Ser Gly Asp Leu Tyr Tyr Ile
530 535 540
Gly Thr
545
<210>21
<211>352
<212>PRT
<213>Nipah virus
<400>21
Met Glu Pro Asp Ile Lys Ser Ile Ser Ser Glu Ser Met Glu Gly Val
1 5 10 15
Ser Asp Phe Ser Pro Ser Ser Trp Glu His Gly Gly Tyr Leu Asp Lys
20 25 30
Val Glu Pro Glu Ile Asp Glu Asn Gly Ser Met Ile Pro Lys Tyr Lys
35 40 45
Ile Tyr Thr Pro Gly Ala Asn Glu Arg Lys Tyr Asn Asn Tyr Met Tyr
50 55 60
Leu Ile Cys Tyr Gly Phe Val Glu Asp Val Glu Arg Thr Pro Glu Thr
65 70 75 80
Gly Lys Arg Lys Lys Ile Arg Thr Ile Ala Ala Tyr Pro Leu Gly Val
85 90 95
Gly Lys Ser Ala Ser His Pro Gln Asp Leu Leu Glu Glu Leu Cys Ser
100 105 110
Leu Lys Val Thr Val Arg Arg Thr Ala Gly Ser Thr Glu Lys Ile Val
115 120 125
Phe Gly Ser Ser Gly Pro Leu Asn His Leu Val Pro Trp Lys Lys Val
130 135 140
Leu Thr Ser Gly Ser Ile Phe Asn Ala Val Lys Val Cys Arg Asn Val
145 150 155 160
Asp Gln Ile Gln Leu Asp Lys His Gln Ala Leu Arg Ile Phe Phe Leu
165 170 175
Ser Ile Thr Lys Leu Asn Asp Ser Gly Ile Tyr Met Ile Pro Arg Thr
180 185 190
Met Leu Glu Phe Arg Arg Asn Asn Ala Ile Ala Phe Asn Leu Leu Val
195 200 205
Tyr Leu Lys Ile Asp Ala Asp Leu Ser Lys Met Gly Ile Gln Gly Ser
210 215 220
Leu Asp Lys Asp Gly Phe Lys Val Ala Ser Phe Met Leu His Leu Gly
225 230 235 240
Asn Phe Val Arg Arg Ala Gly Lys Tyr Tyr Ser Val Asp Tyr Cys Arg
245 250 255
Arg Lys Ile Asp Arg Met Lys Leu Gln Phe Ser Leu Gly Ser Ile Gly
260 265 270
Gly Leu Ser Leu His Ile Lys Ile Asn Gly Val Ile Ser Lys Arg Leu
275 280 285
Phe Ala Gln Met Gly Phe Gln Lys Asn Leu Cys Phe Ser Leu Met Asp
290 295 300
Ile Asn Pro Trp Leu Asn Arg Leu Thr Trp Asn Asn Ser Cys Glu Ile
305 310 315 320
Ser Arg Val Ala Ala Val Leu Gln Pro Ser Ile Pro Arg Glu Phe Met
325 330 335
Ile Tyr Asp Asp Val Phe Ile Asp Asn Thr Gly Arg Ile Leu Lys Gly
340 345 350
<210>22
<211>709
<212>PRT
<213>Nipah virus
<400>22
Met Asp Lys Leu Glu Leu Val Asn Asp Gly Leu Asn Ile Ile Asp Phe
1 5 10 15
Ile Gln Lys Asn Gln Lys Glu Ile Gln Lys Thr Tyr Gly Arg Ser Ser
20 25 30
Ile Gln Gln Pro Ser Ile Lys Asp Gln Thr Lys Ala Trp Glu Asp Phe
35 40 45
Leu Gln Cys Thr Ser Gly Glu Ser Glu Gln Val Glu Gly Gly Met Ser
50 55 60
Lys Asp Asp Gly Asp Val Glu Arg Arg Asn Leu Glu Asp Leu Ser Ser
65 70 75 80
Thr Ser Pro Thr Asp Gly Thr Ile Gly Lys Arg Val Ser Asn Thr Arg
85 90 95
Asp Trp Ala Glu Gly Ser Asp Asp Ile Gln Leu Asp Pro Val Val Thr
100 105 110
Asp Val Val Tyr His Asp His Gly Gly Glu Cys Thr Gly Tyr Gly Phe
115 120 125
Thr Ser Ser Pro Glu Arg Gly Trp Ser Asp Tyr Thr Ser Gly Ala Asn
130 135 140
Asn Gly Asn Val Cys Leu Val Ser Asp Ala Lys Met Leu Ser Tyr Ala
145 150 155 160
Pro Glu Ile Ala Val Ser Lys Glu Asp Arg Glu Thr Asp Leu Val His
165 170 175
Leu Glu Asn Lys Leu Ser Thr Thr Gly Leu Asn Pro Thr Ala Val Pro
180 185 190
Phe Thr Leu Arg Asn Leu Ser Asp Pro Ala Lys Asp Ser Pro Val Ile
195 200 205
Ala Glu His Tyr Tyr Gly Leu Gly Val Lys Glu Gln Asn Val Gly Pro
210 215 220
Gln Thr Ser Arg Asn Val Asn Leu Asp Ser Ile Lys Leu Tyr Thr Ser
225 230 235 240
Asp Asp Glu Glu Ala Asp Gln Leu Glu Phe Glu Asp Glu Phe Ala Gly
245 250 255
Ser Ser Ser Glu Val Ile Val Gly Ile Ser Pro Glu Asp Glu Glu Pro
260 265 270
Ser Ser Val Gly Gly Lys Pro Asn Glu Ser Ile Gly Arg Thr Ile Glu
275 280 285
Gly Gln Ser Ile Arg Asp Asn Leu Gln Ala Lys Asp Asn Lys Ser Thr
290 295 300
Asp Val Pro Gly Ala Gly Pro Lys Asp Ser Ala Val Lys Glu Glu Pro
305 310 315 320
Pro Gln Lys Arg Leu Pro Met Leu Ala Glu Glu Phe Glu Cys Ser Gly
325 330 335
Ser Glu Asp Pro Ile Ile Arg Glu Leu Leu Lys Glu Asn Ser Leu Ile
340 345 350
Asn Cys Gln Gln Gly Lys Asp Ala Gln Pro Pro Tyr His Trp Ser Ile
355 360 365
Glu Arg Ser Ile Ser Pro Asp Lys Thr Glu Ile Val Asn Gly Ala Val
370 375 380
Gln Thr Ala Asp Arg Gln Arg Pro Gly Thr Pro Met Pro Lys Ser Arg
385 390 395 400
Gly Ile Pro Ile Lys Lys Gly Thr Asp Ala Lys Tyr Pro Ser Ala Gly
405 410 415
Thr Glu Asn Val Pro Gly Ser Lys Ser Gly Ala Thr Arg His Val Arg
420 425 430
Gly Ser Pro Pro Tyr Gln Glu Gly Lys Ser Val Asn Ala Glu Asn Val
435 440 445
Gln Leu Asn Ala Ser Thr Ala Val Lys Glu Thr Asp Lys Ser Glu Val
450 455 460
Asn Pro Val Asp Asp Asn Asp Ser Leu Asp Asp Lys Tyr Ile Met Pro
465 470 475 480
Ser Asp Asp Phe Ser Asn Thr Phe Phe Pro His Asp Thr Asp Arg Leu
485 490 495
Asn Tyr His Ala Asp His Leu Gly Asp Tyr Asp Leu Glu Thr Leu Cys
500 505 510
Glu Glu Ser Val Leu Met Gly Val Ile Asn Ser Ile Lys Leu Ile Asn
515 520 525
Leu Asp Met Arg Leu Asn His Ile Glu Glu Gln Val Lys Glu Ile Pro
530 535 540
Lys Ile Ile Asn Lys Leu Glu Ser Ile Asp Arg Val Leu Ala Lys Thr
545 550 555 560
Asn Thr Ala Leu Ser Thr Ile Glu Gly His Leu Val Ser Met Met Ile
565 570 575
Met Ile Pro Gly Lys Gly Lys Gly Glu Arg Lys Gly Lys Asn Asn Pro
580 585 590
Glu Leu Lys Pro Val Ile Gly Arg Asp Ile Leu Glu Gln Gln Ser Leu
595 600 605
Phe Ser Phe Asp Asn Val Lys Asn Phe Arg Asp Gly Ser Leu Thr Asn
610 615 620
Glu Pro Tyr Gly Ala Ala Val Gln Leu Arg Glu Asp Leu Ile Leu Pro
625 630 635 640
Glu Leu Asn Phe Glu Glu Thr Asn Ala Ser Gln Phe Val Pro Met Ala
645 650 655
Asp Asp Ser Ser Arg Asp Val Ile Lys Thr Leu Ile Arg Thr His Ile
660 665 670
Lys Asp Arg Glu Leu Arg Ser Glu Leu Ile Gly Tyr Leu Asn Lys Ala
675 680 685
Glu Asn Asp Glu Glu Ile Gln Glu Ile Ala Asn Thr Val Asn Asp Ile
690 695 700
Ile Asp Gly Asn Ile
705
<210>23
<211>532
<212>PRT
<213>Nipah virus
<400>23
Met Ser Asp Ile Phe Glu Glu Ala Ala Ser Phe Arg Ser Tyr Gln Ser
1 5 10 15
Lys Leu Gly Arg Asp Gly Arg Ala Ser Ala Ala Thr Ala Thr Leu Thr
20 25 30
Thr Lys Ile Arg Ile Phe Val Pro Ala Thr Asn Ser Pro Glu Leu Arg
35 40 45
Trp Glu Leu Thr Leu Phe Ala Leu Asp Val Ile Arg Ser Pro Ser Ala
50 55 60
Ala Glu Ser Met Lys Val Gly Ala Ala Phe Thr Leu Ile Ser Met Tyr
65 70 75 80
Ser Glu Arg Pro Gly Ala Leu Ile Arg Ser Leu Leu Asn Asp Pro Asp
85 90 95
Ile Glu Ala Val Ile Ile Asp Val Gly Ser Met Val Asn Gly Ile Pro
100 105 110
Val Met Glu Arg Arg Gly Asp Lys Ala Gln Glu Glu Met Glu Gly Leu
115 120 125
Met Arg Ile Leu Lys Thr Ala Arg Asp Ser Ser Lys Gly Lys Thr Pro
130 135 140
phe Val Asp Ser Arg Ala Tyr Gly Leu Arg Ile Thr Asp Met Ser Thr
145 150 155 160
Leu Val Ser Ala Val Ile Thr Ile Glu Ala Gln Ile Trp Ile Leu Ile
165 170 175
Ala Lys Ala Val Thr Ala Pro Asp Thr Ala Glu Glu Ser Glu Thr Arg
180 185 190
Arg Trp Ala Lys Tyr Val Gln Gln Lys Arg Val Asn Pro Phe Phe Ala
195 200 205
Leu Thr Gln Gln Trp Leu Thr Glu Met Arg Asn Leu Leu Ser Gln Ser
210 215 220
Leu Ser Val Arg Lys Phe Met Val Glu Ile Leu Ile Glu Val Lys Lys
225 230 235 240
Gly Gly Ser Ala Lys Gly Arg Ala Val Glu Ile Ile Ser Asp Ile Gly
245 250 255
Asn Tyr Val Glu Glu Thr Gly Met Ala Gly Phe Phe Ala Thr Ile Arg
260 265 270
Phe Gly Leu Glu Thr Arg Tyr Pro Ala Leu Ala Leu Asn Glu Phe Gln
275 280 285
Ser Asp Leu Asn Thr Ile Lys Ser Leu Met Leu Leu Tyr Arg Glu Ile
290 295 300
Gly Pro Arg Ala Pro Tyr Met Val Leu Leu Glu Glu Ser Ile Gln Thr
305 310 315 320
Lys Phe Ala Pro Gly Gly Tyr Pro Leu Leu Trp Ser Phe Ala Met Gly
325 330 335
Val Ala Thr Thr Ile Asp Arg Ser Met Gly Ala Leu Asn Ile Asn Arg
340 345 350
Gly Tyr Leu Glu Pro Met Tyr Phe Arg Leu Gly Gln Lys Ser Ala Arg
355 360 365
His His Ala Gly Gly Ile Asp Gln Asn Met Ala Asn Arg Leu Gly Leu
370 375 380
Ser Ser Asp Gln Val Ala Glu Leu Ala Ala Ala Val Gln Glu Thr Ser
385 390 395 400
Ala Gly Arg Gln Glu Ser Asn Val Gln Ala Arg Glu Ala Lys Phe Ala
405 410 415
Ala Gly Gly Val Leu Ile Gly Gly Ser Asp Gln Asp Ile Asp Glu Gly
420 425 430
Glu Glu Pro Ile Glu Gln Ser Gly Arg Gln Ser Val Thr Phe Lys Arg
435 440 445
Glu Met Ser Ile Ser Ser Leu Ala Asn Ser Val Pro Ser Ser Ser Val
450 455 460
Ser Thr Ser Gly Gly Thr Arg Leu Thr Asn Ser Leu Leu Asn Leu Arg
465 470 475 480
Ser Arg Leu Ala Ala Lys Ala Ala Lys Glu Ala Ala Ser Ser Asn Ala
485 490 495
Thr Asp Asp Pro Ala Ile Ser Asn Arg Thr Gln Gly Glu Ser Glu Lys
500 505 510
Lys Asn Asn Gln Asp Leu Lys Pro Ala Gln Asn Asp Leu Asp Phe Val
515 520 525
Arg Ala Asp Val
530
<210>24
<211>709
<212>PRT
<213>Nipah virus
<400>24
Met Asp Lys Leu Glu Leu Val Asn Asp Gly Leu Asn Ile Ile Asp Phe
1 5 10 15
Ile Gln Lys Asn Gln Lys Glu Ile Gln Lys Thr Tyr Gly Arg Ser Ser
20 25 30
Ile Gln Gln Pro Ser Ile Lys Asp Gln Thr Lys Ala Trp Glu Asp Phe
35 40 45
Leu Gln Cys Thr Ser Gly Glu Ser Glu Gln Val Glu Gly Gly Met Ser
50 55 60
Lys Asp Asp Gly Asp Val Glu Arg Arg Asn Leu Glu Asp Leu Ser Ser
65 70 75 80
Thr Ser Pro Thr Asp Gly Thr Ile Gly Lys Arg Val Ser Asn Thr Arg
85 90 95
Asp Trp Ala Glu Gly Ser Asp Asp Ile Gln Leu Asp Pro Val Val Thr
100 105 110
Asp Val Val Tyr His Asp His Gly Gly Glu Cys Thr Gly Tyr Gly Phe
115 120 125
Thr Ser Ser Pro Glu Arg Gly Trp Ser Asp Tyr Thr Ser Gly Ala Asn
130 135 140
Asn Gly Asn Val Cys Leu Val Ser Asp Ala Lys Met Leu Ser Tyr Ala
145 150 155 160
Pro Glu Ile Ala Val Ser Lys Glu Asp Arg Glu Thr Asp Leu Val His
165 170 175
Leu Glu Asn Lys Leu Ser Thr Thr Gly Leu Asn Pro Thr Ala Val Pro
180 185 190
Phe Thr Leu Arg Asn Leu Ser Asp Pro Ala Lys Asp Ser Pro Val Ile
195 200 205
Ala Glu His Tyr Tyr Gly Leu Gly Val Lys Glu Gln Asn Val Gly Pro
210 215 220
Gln Thr Ser Arg Asn Val Asn Leu Asp Ser Ile Lys Leu Tyr Thr Ser
225 230 235 240
Asp Asp Glu Glu Ala Asp Gln Leu Glu Phe Glu Asp Glu Phe Ala Gly
245 250 255
Ser Ser Ser Glu Val Ile Val Gly Ile Ser Pro Glu Asp Glu Glu Pro
260 265 270
Ser Ser Val Gly Gly Lys Pro Asn Glu Ser Ile Gly Arg Thr Ile Glu
275 280 285
Gly Gln Ser Ile Arg Asp Asn Leu Gln Ala Lys Asp Asn Lys Ser Thr
290 295 300
Asp Val Pro Gly Ala Gly Pro Lys Asp Ser Ala Val Lys Glu Glu Pro
305 310 315 320
Pro Gln Lys Arg Leu Pro Met Leu Ala Glu Glu Phe Glu Cys Ser Gly
325 330 335
Ser Glu Asp Pro Ile Ile Arg Glu Leu Leu Lys Glu Asn Ser Leu Ile
340 345 350
Asn Cys Gln Gln Gly Lys Asp Ala Gln Pro Pro Tyr His Trp Ser Ile
355 360 365
Glu Arg Ser Ile Ser Pro Asp Lys Thr Glu Ile Val Asn Gly Ala Val
370 375 380
Gln Thr Ala Asp Arg Gln Arg Pro Gly Thr Pro Met Pro Lys Ser Arg
385 390 395 400
Gly Ile Pro Ile Lys Lys Gly Thr Asp Ala Lys Tyr Pro Ser Ala Gly
405 410 415
Thr Glu Asn Val Pro Gly Ser Lys Ser Gly Ala Thr Arg His Val Arg
420 425 430
Gly Ser Pro Pro Tyr Gln Glu Gly Lys Ser Val Asn Ala Glu Asn Val
435 440 445
Gln Leu Asn Ala Ser Thr Ala Val Lys Glu Thr Asp Lys Ser Glu Val
450 455 460
Asn Pro Val Asp Asp Asn Asp Ser Leu Asp Asp Lys Tyr Ile Met Pro
465 470 475 480
Ser Asp Asp Phe Ser Asn Thr Phe Phe Pro His Asp Thr Asp Arg Leu
485 490 495
Asn Tyr His Ala Asp His Leu Gly Asp Tyr Asp Leu Glu Thr Leu Cys
500 505 510
Glu Glu Ser Val Leu Met Gly Val Ile Asn Ser Ile Lys Leu Ile Asn
515 520 525
Leu Asp Met Arg Leu Asn His Ile Glu Glu Gln Val Lys Glu Ile Pro
530 535 540
Lys Ile Ile Asn Lys Leu Glu Ser Ile Asp Arg Val Leu Ala Lys Thr
545 550 555 560
Asn Thr Ala Leu Ser Thr Ile Glu Gly His Leu Val Ser Met Met Ile
565 570 575
Met Ile Pro Gly Lys Gly Lys Gly Glu Arg Lys Gly Lys Asn Asn Pro
580 585 590
Glu Leu Lys Pro Val Ile Gly Arg Asp Ile Leu Glu Gln Gln Ser Leu
595 600 605
Phe Ser Phe Asp Asn Val Lys Asn Phe Arg Asp Gly Ser Leu Thr Asn
610 615 620
Glu Pro Tyr Gly Ala Ala Val Gln Leu Arg Glu Asp Leu Ile Leu Pro
625 630 635 640
Glu Leu Asn Phe Glu Glu Thr Asn Ala Ser Gln Phe Val Pro Met Ala
645 650 655
Asp Asp Ser Ser Arg Asp Val Ile Lys Thr Leu Ile Arg Thr His Ile
660 665 670
Lys Asp Arg Glu Leu Arg Ser Glu Leu Ile Gly Tyr Leu Asn Lys Ala
675 680 685
Glu Asn Asp Glu Glu Ile Gln Glu Ile Ala Asn Thr Val Asn Asp Ile
690 695 700
Ile Asp Gly Asn Ile
705
<210>25
<211>398
<212>PRT
<213>Nipah virus
<400>25
Met Asp Lys Leu Glu Leu Val Asn Asp Gly Leu Asn Ile Ile Asp Phe
1 5 10 15
Ile Gln Lys Asn Gln Lys Glu Ile Gln Lys Thr Tyr Gly Arg Ser Ser
20 25 30
Ile Gln Gln Pro Ser Ile Lys Asp Gln Thr Lys Ala Trp Glu Asp Phe
35 40 45
Leu Gln Cys Thr Ser Gly Glu Ser Glu Gln Val Glu Gly Gly Met Ser
50 55 60
Lys Asp Asp Gly Asp Val Glu Arg Arg Asn Leu Glu Asp Leu Ser Ser
65 70 75 80
Thr Ser Pro Thr Asp Gly Thr Ile Gly Lys Arg Val Ser Asn Thr Arg
85 90 95
Asp Trp Ala Glu Gly Ser Asp Asp Ile Gln Leu Asp Pro Val Val Thr
100 105 110
Asp Val Val Tyr His Asp His Gly Gly Glu Cys Thr Gly Tyr Gly Phe
115 120 125
Thr Ser Ser Pro Glu Arg Gly Trp Ser Asp Tyr Thr Ser Gly Ala Asn
130 135 140
Asn Gly Asn Val Cys Leu Val Ser Asp Ala Lys Met Leu Ser Tyr Ala
145 150 155 160
Pro Glu Ile Ala Val Ser Lys Glu Asp Arg Glu Thr Asp Leu Val His
165 170 175
Leu Glu Asn Lys Leu Ser Thr Thr Gly Leu Asn Pro Thr Ala Val Pro
180 185 190
Phe Thr Leu Arg Asn Leu Ser Asp Pro Ala Lys Asp Ser Pro Val Ile
195 200 205
Ala Glu His Tyr Tyr Gly Leu Gly Val Lys Glu Gln Asn Val Gly Pro
210 215 220
Gln Thr Ser Arg Asn Val Asn Leu Asp Ser Ile Lys Leu Tyr Thr Ser
225 230 235 240
Asp Asp Glu Glu Ala Asp Gln Leu Glu Phe Glu Asp Glu Phe Ala Gly
245 250 255
Ser Ser Ser Glu Val Ile Val Gly Ile Ser Pro Glu Asp Glu Glu Pro
260 265 270
Ser Ser Val Gly Gly Lys Pro Asn Glu Ser Ile Gly Arg Thr Ile Glu
275 280 285
Gly Gln Ser Ile Arg Asp Asn Leu Gln Ala Lys Asp Asn Lys Ser Thr
290 295 300
Asp Val Pro Gly Ala Gly Pro Lys Asp Ser Ala Val Lys Glu Glu Pro
305 310 315 320
Pro Gln Lys Arg Leu Pro Met Leu Ala Glu Glu Phe Glu Cys Ser Gly
325 330 335
Ser Glu Asp Pro Ile Ile Arg Glu Leu Leu Lys Glu Asn Ser Leu Ile
340 345 350
Asn Cys Gln Gln Gly Lys Asp Ala Gln Pro Pro Tyr His Trp Ser Ile
355 360 365
Glu Arg Ser Ile Ser Pro Asp Lys Thr Glu Ile Val Asn Gly Ala Val
370 375 380
Gln Thr Ala Asp Arg Gln Arg Pro Gly Thr Pro Met Pro Lys
385 390 395
<210>26
<211>166
<212>PRT
<213>Nipah virus
<400>26
Met Met Ala Ser Ile Leu Leu Thr Leu Phe Arg Arg Thr Lys Lys Lys
1 5 10 15
Tyr Arg Arg His Thr Asp Asp Gln Val Phe Asn Asn Pro Ala Ser Lys
20 25 30
Ile Lys Gln Lys Pro Gly Lys Ile Phe Cys Ser Ala Pro Val Glu Asn
35 40 45
Leu Asn Lys Leu Arg Gly Glu Cys Leu Arg Met Met Glu Met Leu Lys
50 55 60
Glu Glu Thr Trp Arg Ile Tyr Pro Val Leu Leu Pro Gln Met Glu Leu
65 70 75 80
Leu Glu Arg Glu Cys Arg Thr Pro Val Thr Gly Gln Lys Val Gln Met
85 90 95
Thr Tyr Asn Trp Thr Gln Trp Leu Gln Thr Leu Tyr Thr Met Ile Met
100 105 110
Glu Glu Asn Val Pro Asp Met Asp Leu Leu Gln Ala Leu Arg Glu Gly
115 120 125
Gly Val Ile Thr His Gln Glu Gln Thr Met Gly Met Tyr Val Leu Tyr
130 135 140
Leu Met Gln Arg Cys Cys Pro Met Leu Pro Lys Leu Gln Phe Leu Lys
145 150 155 160
Lys Ile Gly Lys Leu Ile
165
<210>27
<211>352
<212>PRT
<213>Nipah virus
<400>27
Met Glu Pro Asp Ile Lys Ser Ile Ser Ser Glu Ser Met Glu Gly Val
1 5 10 15
Ser Asp Phe Ser Pro Ser Ser Trp Glu His Gly Gly Tyr Leu Asp Lys
20 25 30
Val Glu Pro Glu Ile Asp Glu Asn Gly Ser Met Ile Pro Lys Tyr Lys
35 40 45
Ile Tyr Thr Pro Gly Ala Asn Glu Arg Lys Tyr Asn Asn Tyr Met Tyr
50 55 60
Leu Ile Cys Tyr Gly Phe Val Glu Asp Val Glu Arg Thr Pro Glu Thr
65 70 75 80
Gly Lys Arg Lys Lys Ile Arg Thr Ile Ala Ala Tyr Pro Leu Gly Val
85 90 95
Gly Lys Ser Ala Ser His Pro Gln Asp Leu Leu Glu Glu Leu Cys Ser
100 105 110
Leu Lys Val Thr Val Arg Arg Thr Ala Gly Ser Thr Glu Lys Ile Val
115 120 125
Phe Gly Ser Ser Gly Pro Leu Asn His Leu Val Pro Trp Lys Lys Val
130 135 140
Leu Thr Ser Gly Ser Ile Phe Asn Ala Val Lys Val Cys Arg Asn Val
145 150 155 160
Asp Gln Ile Gln Leu Asp Lys His Gln Ala Leu Arg Ile Phe Phe Leu
165 170 175
Ser Ile Thr Lys Leu Asn Asp Ser Gly Ile Tyr Met Ile Pro Arg Thr
180 185 190
Met Leu Glu Phe Arg Arg Asn Asn Ala Ile Ala Phe Asn Leu Leu Val
195 200 205
Tyr Leu Lys Ile Asp Ala Asp Leu Ser Lys Met Gly Ile Gln Gly Ser
210 215 220
Leu Asp Lys Asp Gly Phe Lys Val Ala Ser Phe Met Leu His Leu Gly
225 230 235 240
Asn Phe Val Arg Arg Ala Gly Lys Tyr Tyr Ser Val Asp Tyr Cys Arg
245 250 255
Arg Lys Ile Asp Arg Met Lys Leu Gln Phe Ser Leu Gly Ser Ile Gly
260 265 270
Gly Leu Ser Leu His Ile Lys Ile Asn Gly Val Ile Ser Lys Arg Leu
275 280 285
Phe Ala Gln Met Gly Phe Gln Lys Asn Leu Cys Phe Ser Leu Met Asp
290 295 300
Ile Asn Pro Trp Leu Asn Arg Leu Thr Trp Asn Asn Ser Cys Glu Ile
305 310 315 320
Ser Arg Val Ala Ala Val Leu Gln Pro Ser Ile Pro Arg Glu Phe Met
325 330 335
Ile Tyr Asp Asp Val Phe Ile Asp Asn Thr Gly Arg Ile Leu Lys Gly
340 345 350
<210>28
<211>546
<212>PRT
<213>Nipah virus
<400>28
Met Val Val Ile Leu Asp Lys Arg Cys Tyr Cys Asn Leu Leu Ile Leu
1 5 10 15
Ile Leu Met Ile Ser Glu Cys Ser Val Gly Ile Leu His Tyr Glu Lys
20 25 30
Leu Ser Lys Ile Gly Leu Val Lys Gly Val Thr Arg Lys Tyr Lys Ile
35 40 45
Lys Ser Asn Pro Leu Thr Lys Asp Ile Val Ile Lys Met Ile Pro Asn
50 55 60
Val Ser Asn Met Ser Gln Cys Thr Gly Ser Val Met Glu Asn Tyr Lys
65 70 75 80
Thr Arg Leu Asn Gly Ile Leu Thr Pro Ile Lys Gly Ala Leu Glu Ile
85 90 95
Tyr Lys Asn Asn Thr His Asp Leu Val Gly Asp Val Arg Leu Ala Gly
100 105 110
Val Ile Met Ala Gly Val Ala Ile Gly Ile Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Val Ala Leu Tyr Glu Ala Met Lys Asn Ala Asp Asn Ile
130 135 140
Asn Lys Leu Lys Ser Ser Ile Glu Ser Thr Asn Glu Ala Val Val Lys
145 150 155 160
Leu Gln Glu Thr Ala Glu Lys Thr Val Tyr Val Leu Thr Ala Leu Gln
165 170 175
Asp Tyr Ile Asn Thr Asn Leu Val Pro Thr Ile Asp Lys Ile Ser Cys
180 185 190
Lys Gln Thr Glu Leu Ser Leu Asp Leu Ala Leu Ser Lys Tyr Leu Ser
195 200 205
Asp Leu Leu Phe Val Phe Gly Pro Asn Leu Gln Asp Pro Val Ser Asn
210 215 220
Ser Met Thr Ile Gln Ala Ile Ser Gln Ala Phe Gly Gly Asn Tyr Glu
225 230 235 240
Thr Leu Leu Arg Thr Leu Gly Tyr Ala Thr Glu Asp Phe Asp Asp Leu
245 250 255
Leu Glu Ser Asp Ser Ile Thr Gly Gln Ile Ile Tyr Val Asp Leu Ser
260 265 270
Ser Tyr Tyr Ile Ile Val Arg Val Tyr Phe Pro Ile Leu Thr Glu Ile
275 280 285
Gln Gln Ala Tyr Ile Gln Glu Leu Leu Pro Val Ser Phe Asn Asn Asp
290 295 300
Asn Ser Glu Trp Ile Ser Ile Val Pro Asn Phe Ile Leu Val Arg Asn
305 310 315 320
Thr Leu Ile Ser Asn Ile Glu Ile Gly Phe Cys Leu Ile Thr Lys Arg
325 330 335
Ser Val Ile Cys Asn Gln Asp Tyr Ala Thr Pro Met Thr Asn Asn Met
340 345 350
Arg Glu Cys Leu Thr Gly Ser Thr Glu Lys Cys Pro Arg Glu Leu Val
355 360 365
Val Ser Ser His Val Pro Arg Phe Ala Leu Ser Asn Gly Val Leu Phe
370 375 380
Ala Asn Cys Ile Ser Val Thr Cys Gln Cys Gln Thr Thr Gly Arg Ala
385 390 395 400
Ile Ser Gln Ser Gly Glu Gln Thr Leu Leu Met Ile Asp Asn Thr Thr
405 410 415
Cys Pro Thr Ala Val Leu Gly Asn Val Ile Ile Ser Leu Gly Lys Tyr
420 425 430
Leu Gly Ser Val Asn Tyr Asn Ser Glu Gly Ile Ala Ile Gly Pro Pro
435 440 445
Val Phe Thr Asp Lys Val Asp Ile Ser Ser Gln Ile Ser Ser Met Asn
450 455 460
Gln Ser Leu Gln Gln Ser Lys Asp Tyr Ile Lys Glu Ala Gln Arg Leu
465 470 475 480
Leu Asp Thr Val Asn Pro Ser Leu Ile Ser Met Leu Ser Met Ile Ile
485 490 495
Leu Tyr Val Leu Ser Ile Ala Ser Leu Cys Ile Gly Leu Ile Thr Phe
500 505 510
Ile Ser Phe Ile Ile Val Glu Lys Lys Arg Asn Thr Tyr Ser Arg Leu
515 520 525
Glu Asp Arg Arg Val Arg Pro Thr Ser Ser Gly Asp Leu Tyr Tyr Ile
530 535 540
Gly Thr
545
<210>29
<211>602
<212>PRT
<213>Nipah virus
<400>29
Met Pro Ala Glu Asn Lys Lys Val Arg Phe Glu Asn Thr Thr Ser Asp
1 5 10 15
Lys Gly Lys Ile Pro Ser Lys Val Ile Lys Ser Tyr Tyr Gly Thr Met
20 25 30
Asp Ile Lys Lys Ile Asn Glu Gly Leu Leu Asp Ser Lys Ile Leu Ser
35 40 45
Ala Phe Asn Thr Val Ile Ala Leu Leu Gly Ser Ile Val Ile Ile Val
50 55 60
Met Asn Ile Met Ile Ile Gln Asn Tyr Thr Arg Ser Thr Asp Asn Gln
65 70 75 80
Ala Val Ile Lys Asp Ala Leu Gln Gly Ile Gln Gln Gln Ile Lys Gly
85 90 95
Leu Ala Asp Lys Ile Gly Thr Glu Ile Gly Pro Lys Val Ser Leu Ile
100 105 110
Asp Thr Ser Ser Thr Ile Thr Ile Pro Ala Asn Ile Gly Leu Leu Gly
115 120 125
Ser Lys Ile Ser Gln Ser Thr Ala Ser Ile Asn Glu Asn Val Asn Glu
130 135 140
Lys Cys Lys Phe Thr Leu Pro Pro Leu Lys Ile His Glu Cys Asn Ile
145 150 155 160
Ser Cys Pro Asn Pro Leu Pro Phe Arg Glu Tyr Arg Pro Gln Thr Glu
165 170 175
Gly Val Ser Asn Leu Val Gly Leu Pro Asn Asn Ile Cys Leu Gln Lys
180 185 190
Thr Ser Asn Gln Ile Leu Lys Pro Lys Leu Ile Ser Tyr Thr Leu Pro
195 200 205
Val Val Gly Gln Ser Gly Thr Cys Ile Thr Asp Pro Leu Leu Ala Met
210 215 220
Asp Glu Gly Tyr Phe Ala Tyr Ser His Leu Glu Arg Ile Gly Ser Cys
225 230 235 240
Ser Arg Gly Val Ser Lys Gln Arg Ile Ile Gly Val Gly Glu Val Leu
245 250 255
Asp Arg Gly Asp Glu Val Pro Ser Leu Phe Met Thr Asn Val Trp Thr
260 265 270
Pro Pro Asn Pro Asn Thr Val Tyr His Cys Ser Ala Val Tyr Asn Asn
275 280 285
Glu Phe Tyr Tyr Val Leu Cys Ala Val Ser Thr Val Gly Asp Pro Ile
290 295 300
Leu Asn Ser Thr Tyr Trp Ser Gly Ser Leu Met Met Thr Arg Leu Ala
305 310 315 320
Val Lys Pro Lys Ser Asn Gly Gly Gly Tyr Asn Gln His Gln Leu Ala
325 330 335
Leu Arg Ser Ile Glu Lys Gly Arg Tyr Asp Lys Val Met Pro Tyr Gly
340 345 350
Pro Ser Gly Ile Lys Gln Gly Asp Thr Leu Tyr Phe Pro Ala Val Gly
355 360 365
Phe Leu Val Arg Thr Glu Phe Lys Tyr Asn Asp Ser Asn Cys Pro Ile
370 375 380
Thr Lys Cys Gln Tyr Ser Lys Pro Glu Asn Cys Arg Leu Ser Met Gly
385 390 395 400
Ile Arg Pro Asn Ser His Tyr Ile Leu Arg Ser Gly Leu Leu Lys Tyr
405 410 415
Asn Leu Ser Asp Gly Glu Asn Pro Lys Val Val Phe Ile Glu Ile Ser
420 425 430
Asp Gln Arg Leu Ser Ile Gly Ser Pro Ser Lys Ile Tyr Asp Ser Leu
435 440 445
Gly Gln Pro Val Phe Tyr Gln Ala Ser Phe Ser Trp Asp Thr Met Ile
450 455 460
Lys Phe Gly Asp Val Leu Thr Val Asn Pro Leu Val Val Asn Trp Arg
465 470 475 480
Asn Asn Thr Val Ile Ser Arg Pro Gly Gln Ser Gln Cys Pro Arg Phe
485 490 495
Asn Thr Cys Pro Glu Ile Cys Trp Glu Gly Val Tyr Asn Asp Ala Phe
500 505 510
Leu Ile Asp Arg Ile Asn Trp Ile Ser Ala Gly Val Phe Leu Asp Ser
515 520 525
Asn Gln Thr Ala Glu Asn Pro Val Phe Thr Val Phe Lys Asp Asn Glu
530 535 540
Ile Leu Tyr Arg Ala Gln Leu Ala Ser Glu Asp Thr Asn Ala Gln Lys
545 550 555 560
Thr Ile Thr Asn Cys Phe Leu Leu Lys Asn Lys Ile Trp Cys Ile Ser
565 570 575
Leu Val Glu Ile Tyr Asp Thr Gly Asp Asn Val Ile Arg Pro Lys Leu
580 585 590
Phe Ala Val Lys Ile Pro Glu Gln Cys Thr
595 600
<210>30
<211>2244
<212>PRT
<213>Nipah virus
<400>30
Met Ala Asp Glu Leu Ser Ile Ser Asp Ile Ile Tyr Pro Glu Cys His
1 5 10 15
Leu Asp Ser Pro Ile Val Ser Gly Lys Leu Ile Ser Ala Ile Glu Tyr
20 25 30
Ala Gln Leu Arg His Asn Gln Pro Ser Asp Asp Lys Arg Leu Ser Glu
35 40 45
Asn Ile Arg Leu Asn Leu His Gly Lys Arg Lys Ser Leu Tyr Ile Leu
50 55 60
Arg Gln Ser Lys Gln Gly Asp Tyr Ile Arg Asn Asn Ile Lys Asn Leu
65 70 75 80
Lys Glu Phe Met His Ile Ala Tyr Pro Glu Cys Asn Asn Ile Leu Phe
85 90 95
Ser Ile Thr Ser Gln Gly Met Thr Ser Lys Leu Asp Asn Ile Met Lys
100 105 110
Lys Ser Phe Lys Ala Tyr Asn Ile Ile Ser Lys Lys Val Ile Gly Met
115 120 125
Leu Gln Asn Ile Thr Arg Asn Leu Ile Thr Gln Asp Arg Arg Asp Glu
130 135 140
Ile Ile Asn Ile His Glu Cys Arg Arg Leu Gly Asp Leu Gly Lys Asn
145 150 155 160
Met Ser Gln Ser Lys Trp Tyr Glu Cys Phe Leu Phe Trp Phe Thr Ile
165 170 175
Lys Thr Glu Met Arg Ala Val Ile Lys Asn Ser Gln Lys Pro Lys Phe
180 185 190
Arg Ser Asp Ser Cys Ile Ile His Met Arg Asp Lys Ser Thr Glu Ile
195 200 205
Ile Leu Asn Pro Asn Leu Ile Cys Ile Phe Lys Ser Asp Lys Thr Gly
210 215 220
Lys Lys Cys Tyr Tyr Leu Thr Pro Glu Met Val Leu Met Tyr Cys Asp
225 230 235 240
Val Leu Glu Gly Arg Met Met Met Glu Thr Thr Val Lys Ser Asp Ile
245 250 255
Lys Tyr Gln Pro Leu Ile Ser Arg Ser Asn Ala Leu Trp Gly Leu Ile
260 265 270
Asp Pro Leu Phe Pro Val Met Gly Asn Arg Ile Tyr Asn Ile Val Ser
275 280 285
Met Ile Glu Pro Leu Val Leu Ala Leu Leu Gln Leu Lys Asp Glu Ala
290 295 300
Arg Ile Leu Arg Gly Ala Phe Leu His His Cys Ile Lys Glu Met His
305 310 315 320
Gln Glu Leu Ser Glu Cys Gly Phe Thr Asp Gln Lys Ile Arg Ser Met
325 330 335
Phe Ile Asp Asp Leu Leu Ser Ile Leu Asn Ile Asp Asn Ile His Leu
340 345 350
Leu Ala Glu Phe Phe Ser Phe Phe Arg Thr Phe Gly His Pro Ile Leu
355 360 365
Glu Ala Lys Val Ala Ala Glu Lys Val Arg Glu His Met Leu Ala Asp
370 375 380
Lys Val Leu Glu Tyr Ala Pro Ile Met Lys Ala His Ala Ile Phe Cys
385 390 395 400
Gly Thr Ile Ile Asn Gly Tyr Arg Asp Arg His Gly Gly Ala Trp Pro
405 410 415
Pro Leu Tyr Leu Pro Ala His Ala Ser Lys His Ile Ile Arg Leu Lys
420 425 430
Asn Ser Gly Glu Ser Leu Thr Ile Asp Asp Cys Val Lys Asn Trp Glu
435 440 445
Ser Phe Cys Gly Ile Gln Phe Asp Cys Phe Met Glu Leu Lys Leu Asp
450 455 460
Ser Asp Leu Ser Met Tyr Met Lys Asp Lys Ala Leu Ser Pro Ile Lys
465 470 475 480
Asp Glu Trp Asp Ser Val Tyr Pro Arg Glu Val Leu Ser Tyr Thr Pro
485 490 495
Pro Lys Ser Thr Glu Pro Arg Arg Leu Val Asp Val Phe Val Asn Asp
500 505 510
Glu Asn Phe Asp Pro Tyr Asn Met Leu Glu Tyr Val Leu Ser Gly Ala
515 520 525
Tyr Leu Glu Asp Glu Gln Phe Asn Val Ser Tyr Ser Leu Lys Glu Lys
530 535 540
Glu Thr Lys Gln Ala Gly Arg Leu Phe Ala Lys Met Thr Tyr Lys Met
545 550 555 560
Arg Ala Cys Gln Val Ile Ala Glu Ala Leu Ile Ala Ser Gly Val Gly
565 570 575
Lys Tyr Phe Lys Glu Asn Gly Met Val Lys Asp Glu His Glu Leu Leu
580 585 590
Lys Thr Leu Phe Gln Leu Ser Ile Ser Ser Val Pro Arg Gly Asn Ser
595 600 605
Gln Gly Asn Asp Pro Gln Ser Ile Asn Asn Ile Glu Arg Asp Phe Gln
610 615 620
Tyr Phe Lys Gly Val Thr Thr Asn Val Lys Asp Lys Lys Asn Asn Ser
625 630 635 640
Phe Asn Lys Val Lys Ser Ala Leu Asn Asn Pro Cys Gln Ala Asp Gly
645 650 655
Val His His Asn Met Ser Pro Asn Thr Arg Asn Arg Tyr Lys Cys Ser
660 665 670
Asn Thr Ser Lys Ser Phe Leu Asp Tyr His Thr Glu Phe Asn Pro His
675 680 685
Asn His Tyr Lys Ser Asp Asn Thr Glu Ala Ala Val Leu Ser Arg Tyr
690 695 700
Glu Asp Asn Thr Gly Thr Lys Phe Asp Thr Val Ser Ala Phe Leu Thr
705 710 715 720
Thr Asp Leu Lys Lys Phe Cys Leu Asn Trp Arg Tyr Glu Ser Met Ala
725 730 735
Ile Phe Ala Glu Arg Leu Asp Glu Ile Tyr Gly Leu Pro Gly Phe Phe
740 745 750
Asn Trp Met His Lys Arg Leu Glu Arg Ser Val Ile Tyr Val Ala Asp
755 760 765
Pro Asn Cys Pro Pro Asn Ile Asp Lys His Met Glu Leu Glu Lys Thr
770 775 780
Pro Glu Asp Asp Ile Phe Ile His Tyr Pro Lys Gly Gly Ile Glu Gly
785 790 795 800
Tyr Ser Gln Lys Thr Trp Thr Ile Ala Thr Ile Pro Phe Leu Phe Leu
805 810 815
Ser Ala Tyr Glu Thr Asn Thr Arg Ile Ala Ala Ile Val Gln Gly Asp
820 825 830
Asn Glu Ser Ile Ala Ile Thr Gln Lys Val His Pro Asn Leu Pro Tyr
835 840 845
Lys Val Lys Lys Glu Ile Cys Ala Lys Gln Ala Gln Leu Tyr Phe Glu
850 855 860
Arg Leu Arg Met Asn Leu Arg Ala Leu Gly His Asn Leu Lys Ala Thr
865 870 875 880
Glu Thr Ile Ile Ser Thr His Leu Phe Ile Tyr Ser Lys Lys Ile His
885 890 895
Tyr Asp Gly Ala Val Leu Ser Gln Ala Leu Lys Ser Met Ser Arg Cys
900 905 910
Cys Phe Trp Ser Glu Thr Leu Val Asp Glu Thr Arg Ser Ala Cys Ser
915 920 925
Asn Ile Ser Thr Thr Ile Ala Lys Ala Ile Glu Asn Gly Leu Ser Arg
930 935 940
Asn Val Gly Tyr Cys Ile Asn Ile Leu Lys Val Ile Gln Gln Leu Leu
945 950 955 960
Ile Ser Thr Glu Phe Ser Ile Asn Glu Thr Leu Thr Leu Asp Val Thr
965 970 975
Ser Pro Ile Ser Asn Asn Leu Asp Trp Leu Ile Thr Ala Ala Leu Ile
980 985 990
Pro Ala Pro Ile Gly Gly Phe Asn Tyr Leu Asn Leu Ser Arg Ile Phe
995 1000 1005
Val Arg Asn Ile Gly Asp Pro Val Thr Ala Ser Leu Ala Asp Leu
1010 1015 1020
Lys Arg Met Ile Asp His Ser Ile Met Thr Glu Ser Val Leu Gln
1025 1030 1035
Lys Val Met Asn Gln Glu Pro Gly Asp Ala Ser Phe Leu Asp Trp
1040 1045 1050
Ala Ser Asp Pro Tyr Ser Gly Asn Leu Pro Asp Ser Gln Ser Ile
1055 1060 1065
Thr Lys Thr Ile Lys Asn Ile Thr Ala Arg Thr Ile Leu Arg Asn
1070 1075 1080
Ser Pro Asn Pro Met Leu Lys Gly Leu Phe His Asp Lys Ser Phe
1085 1090 1095
Asp Glu Asp Leu Glu Leu Ala Ser Phe Leu Met Asp Arg Arg Val
1100 1105 1110
Ile Leu Pro Arg Ala Ala His Glu Ile Leu Asp Asn Ser Leu Thr
1115 1120 1125
Gly Ala Arg Glu Glu Ile Ala Gly Leu Leu Asp Thr Thr Lys Gly
1130 1135 1140
Leu Ile Arg Ser Gly Leu Arg Lys Ser Gly Leu Gln Pro Lys Leu
1145 1150 1155
Val Ser Arg Leu Ser His His Asp Tyr Asn Gln Phe Leu Ile Leu
1160 1165 1170
Asn Lys Leu Leu Ser Asn Arg Arg Gln Asn Asp Leu Ile Ser Ser
1175 1180 1185
Asn Thr Cys Ser Val Asp Leu Ala Arg Ala Leu Arg Ser His Met
1190 1195 1200
Trp Arg Glu Leu Ala Leu Gly Arg Val Ile Tyr Gly Leu Glu Val
1205 1210 1215
Pro Asp Ala Leu Glu Ala Met Val Gly Arg Tyr Ile Thr Gly Ser
1220 1225 1230
Leu Glu Cys Gln Ile Cys Glu Gln Gly Asn Thr Met Tyr Gly Trp
1235 1240 1245
Phe Phe Val Pro Arg Asp Ser Gln Leu Asp Gln Val Asp Arg Glu
1250 1255 1260
His Ser Ser Ile Arg Val Pro Tyr Val Gly Ser Ser Thr Asp Glu
1265 1270 1275
Arg Ser Asp Ile Lys Leu Gly Asn Val Lys Arg Pro Thr Lys Ala
1280 1285 1290
Leu Arg Ser Ala Ile Arg Ile Ala Thr Val Tyr Thr Trp Ala Tyr
1295 1300 1305
Gly Asp Asn Glu Glu Cys Trp Tyr Glu Ala Trp Tyr Leu Ala Ser
1310 1315 1320
Gln Arg Val Asn Ile Asp Leu Asp Val Leu Lys Ala Ile Thr Pro
1325 1330 1335
Val Ser Thr Ser Asn Asn Leu Ser His Arg Leu Arg Asp Lys Ser
1340 1345 1350
Thr Gln Phe Lys Phe Ala Gly Ser Val Leu Asn Arg Val Ser Arg
1355 1360 1365
Tyr Val Asn Ile Ser Asn Asp Asn Leu Asp Phe Arg Ile Glu Gly
1370 1375 1380
Glu Lys Val Asp Thr Asn Leu Ile Tyr Gln Gln Ala Met Leu Leu
1385 1390 1395
Gly Leu Ser Val Leu Glu Gly Lys Phe Arg Leu Arg Leu Glu Thr
1400 1405 1410
Asp Asp Tyr Asn Gly Ile Tyr His Leu His Val Lys Asp Asn Cys
1415 1420 1425
Cys Val Lys Glu Val Ala Asp Val Gly Gln Val Asp Ala Glu Leu
1430 1435 1440
Pro Ile Pro Glu Tyr Thr Glu Val Asp Asn Asn His Leu Ile Tyr
1445 1450 1455
Asp Pro Asp Pro Val Ser Glu Ile Asp Cys Ser Arg Leu Ser Asn
1460 1465 1470
Gln Glu Ser Lys Ser Arg Glu Leu Asp Phe Pro Leu Trp Ser Thr
1475 1480 1485
Glu Glu Leu His Asp Val Leu Ala Lys Thr Val Ala Gln Thr Val
1490 1495 1500
Leu Glu Ile Ile Thr Lys Ala Asp Lys Asp Val Leu Lys Gln His
1505 1510 1515
Leu Ala Ile Asp Ser Asp Asp Asn Ile Asn Ser Leu Ile Thr Glu
1520 1525 1530
Phe Leu Ile Val Asp Pro Glu Leu Phe Ala Leu Tyr Leu Gly Gln
1535 1540 1545
Ser Ile Ser Ile Lys Trp Ala Phe Glu Ile His His Arg Arg Pro
1550 1555 1560
Arg Gly Arg His Thr Met Val Asp Leu Leu Ser Asp Leu Val Ser
1565 1570 1575
Asn Thr Ser Lys His Thr Tyr Lys Val Leu Ser Asn Ala Leu Ser
1580 1585 1590
His Pro Arg Val Phe Lys Arg Phe Val Asn Cys Gly Leu Leu Leu
1595 1600 1605
Pro Thr Gln Gly Pro Tyr Leu His Gln Gln Asp Phe Glu Lys Leu
1610 1615 1620
Ser Gln Asn Leu Leu Val Thr Ser Tyr Met Ile Tyr Leu Met Asn
1625 1630 1635
Trp Cys Asp Phe Lys Lys Ser Pro Phe Leu Ile Ala Glu Gln Asp
1640 1645 1650
Glu Thr Val Ile Ser Leu Arg Glu Asp Ile Ile Thr Ser Lys His
1655 1660 1665
Leu Cys Val Ile Ile Asp Leu Tyr Ala Asn His His Lys Pro Pro
1670 1675 1680
Trp Ile Ile Asp Leu Asn Pro Gln Glu Lys Ile Cys Val Leu Arg
1685 1690 1695
Asp Phe Ile Ser Lys Ser Arg His Val Asp Thr Ser Ser Arg Ser
1700 1705 1710
Trp Asn Thr Ser Asp Leu Asp Phe Val Ile Phe Tyr Ala Ser Leu
1715 1720 1725
Thr Tyr Leu Arg Arg Gly Ile Ile Lys Gln Leu Arg Ile Arg Gln
1730 1735 1740
Val Thr Glu Val Ile Asp Thr Thr Thr Met Leu Arg Asp Asn Ile
1745 1750 1755
Ile Val Glu Asn Pro Pro Ile Lys Thr Gly Val Leu Asp Ile Arg
1760 1765 1770
Gly Cys Ile Ile Tyr Asn Leu Glu Glu Ile Leu Ser Met Asn Thr
1775 1780 1785
Lys Ser Ala Ser Lys Lys Ile Phe Asn Leu Asn Ser Arg Pro Ser
1790 1795 1800
Val Glu Asn His Lys Tyr Arg Arg Ile Gly Leu Asn Ser Ser Ser
18051 810 1815
Cys Tyr Lys Ala Leu Asn Leu Ser Pro Leu Ile Gln Arg Tyr Leu
1820 1825 1830
Pro Ser Gly Ala Gln Arg Leu Phe Ile Gly Glu Gly Ser Gly Ser
1835 1840 1845
Met Met Leu Leu Tyr Gln Ser Thr Leu Gly Gln Ser Ile Ser Phe
1850 1855 1860
Tyr Asn Ser Gly Ile Asp Gly Asp Tyr Ile Pro Gly Gln Arg Glu
1865 1870 1875
Leu Lys Leu Phe Pro Ser Glu Tyr Ser Ile Ala Glu Glu Asp Pro
1880 1885 1890
Ser Leu Thr Gly Lys Leu Lys Gly Leu Val Val Pro Leu Phe Asn
1895 1900 1905
Gly Arg Pro Glu Thr Thr Trp Ile Gly Asn Leu Asp Ser Tyr Glu
1910 1915 1920
Tyr Ile Ile Asn Arg Thr Ala Gly Arg Ser Ile Gly Leu Val His
1925 1930 1935
Ser Asp Met Glu Ser Gly Ile Asp Lys Asn Val Glu Glu Ile Leu
1940 1945 1950
Val Glu His Ser His Leu Ile Ser Ile Ala Ile Asn Val Met Met
1955 1960 1965
Glu Asp Gly Leu Leu Val Ser Lys Ile Ala Tyr Thr Pro Gly Phe
1970 1975 1980
Pro Ile Ser Arg Leu Phe Asn Met Tyr Arg Ser Tyr Phe Gly Leu
1985 1990 1995
Val Leu Val Cys Phe Pro Val Tyr Ser Asn Pro Asp Ser Thr Glu
2000 2005 2010
Val Tyr Leu Leu Cys Leu Gln Lys Thr Val Lys Thr Ile Val Pro
2015 2020 2025
Pro Gln Lys Val Leu Glu His Ser Asn Leu His Asp Glu Val Asn
2030 2035 2040
Asp Gln Gly Ile Thr Ser Val Ile Phe Lys Ile Lys Asn Ser Gln
2045 2050 2055
Ser Lys Gln Phe His Asp Asp Leu Lys Lys Tyr Tyr Gln Ile Asp
2060 2065 2070
Gln Pro Phe Phe Val Pro Thr Lys Ile Thr Ser Asp Glu Gln Val
2075 2080 2085
Leu Leu Gln Ala Gly Leu Lys Leu Asn Gly Pro Glu Ile Leu Lys
2090 2095 2100
Ser Glu Ile Ser Tyr Asp Ile Gly Ser Asp Ile Asn Thr Leu Arg
2105 2110 2115
Asp Thr Ile Ile Ile Met Leu Asn Glu Ala Met Asn Tyr Phe Asp
2120 2125 2130
Asp Asn Arg Ser Pro Ser His His Leu Glu Pro Tyr Pro Val Leu
2135 2140 2145
Glu Arg Thr Arg Ile Lys Thr Ile Met Asn Cys Val Thr Lys Lys
2150 2155 2160
Val Ile Val Tyr Ser Leu Ile Lys Phe Lys Asp Thr Lys Ser Ser
2165 2170 2175
Glu Leu Tyr His Ile Lys Asn Asn Ile Arg Arg Lys Val Leu Ile
2180 2185 2190
Leu Asp Phe Arg Ser Lys Leu Met Thr Lys Thr Leu Pro Lys Gly
2195 2200 2205
Met Gln Glu Arg Arg Glu Lys Asn Gly Phe Lys Glu Val Trp Ile
2210 2215 2220
Val Asp Leu Ser Asn Arg Glu Val Lys Ile Trp Trp Lys Ile Ile
2225 2230 2235
Gly Tyr Ile Ser Ile Ile
2240
<210>31
<211>532
<212>PRT
<213>Nipah virus
<400>3l
Met Ser Asp Ile Phe Glu Glu Ala Ala Ser Phe Arg Ser Tyr Gln Ser
1 5 10 15
Lys Leu Gly Arg Asp Gly Arg Ala Ser Ala Ala Thr Ala Thr Leu Thr
20 25 30
Thr Lys Ile Arg Ile Phe Val Pro Ala Thr Asn Ser Pro Glu Leu Arg
35 40 45
Trp Glu Leu Thr Leu Phe Ala Leu Asp Val Ile Arg Ser Pro Ser Ala
50 55 60
Ala Glu Ser Met Lys Val Gly Ala Ala Phe Thr Leu Ile Ser Met Tyr
65 70 75 80
Ser Glu Arg Pro Gly Ala Leu Ile Arg Ser Leu Leu Asn Asp Pro Asp
85 90 95
Ile Glu Ala Val Ile Ile Asp Val Gly Ser Met Val Asn Gly Ile Pro
100 105 110
Val Met Glu Arg Arg Gly Asp Lys Ala Gln Glu Glu Met Glu Gly Leu
115 120 125
Met Arg Ile Leu Lys Thr Ala Arg Asp Ser Ser Lys Gly Lys Thr Pro
130 135 140
Phe Val Asp Ser Arg Ala Tyr Gly Leu Arg Ile Thr Asp Met Ser Thr
145 150 155 160
Leu Val Ser Ala Val Ile Thr Ile Glu Ala Gln Ile Trp Ile Leu Ile
165 170 175
Ala Lys Ala Val Thr Ala Pro Asp Thr Ala Glu Glu Ser Glu Thr Arg
180 185 190
Arg Trp Ala Lys Tyr Val Gln Gln Lys Arg Val Asn Pro Phe Phe Ala
195 200 205
Leu Thr Gln Gln Trp Leu Thr Glu Met Arg Asn Leu Leu Ser Gln Ser
210 215 220
Leu Ser Val Arg Lys Phe Met Val Glu Ile Leu Ile Glu Val Lys Lys
225 230 235 240
Gly Gly Ser Ala Lys Gly Arg Ala Val Glu Ile Ile Ser Asp Ile Gly
245 250 255
Asn Tyr Val Glu Glu Thr Gly Met Ala Gly Phe Phe Ala Thr Ile Arg
260 265 270
Phe Gly Leu Glu Thr Arg Tyr Pro Ala Leu Ala Leu Asn Glu Phe Gln
275 280 285
Ser Asp Leu Asn Thr Ile Lys Ser Leu Met Leu Leu Tyr Arg Glu Ile
290 295 300
Gly Pro Arg Ala Pro Tyr Met Val Leu Leu Glu Glu Ser Ile Gln Thr
305 310 315 320
Lys Phe Ala Pro Gly Gly Tyr Pro Leu Leu Trp Ser Phe Ala Met Gly
325 330 335
Val Ala Thr Thr Ile Asp Arg Ser Met Gly Ala Leu Asn Ile Asn Arg
340 345 350
Gly Tyr Leu Glu Pro Met Tyr Phe Arg Leu Gly Gln Lys Ser Ala Arg
355 360 365
His His Ala Gly Gly Ile Asp Gln Asn Met Ala Asn Arg Leu Gly Leu
370 375 380
Ser Ser Asp Gln Val Ala Glu Leu Ala Ala Ala Val Gln Glu Thr Ser
385 390 395 400
Ala Gly Arg Gln Glu Ser Asn Val Gln Ala Arg Glu Ala Lys Phe Ala
405 410 415
Ala Gly Gly Val Leu Ile Gly Gly Ser Asp Gln Asp Ile Asp Glu Gly
4204 25 430
Glu Glu Pro Ile Glu Gln Ser Gly Arg Gln Ser Val Thr Phe Lys Arg
435 440 445
Glu Met Ser Ile Ser Ser Leu Ala Asn Ser Val Pro Ser Ser Ser Val
450 455 460
Ser Thr Ser Gly Gly Thr Arg Leu Thr Asn Ser Leu Leu Asn Leu Arg
465 470 475 480
Ser Arg Leu Ala Ala Lys Ala Ala Lys Glu Ala Ala Ser Ser Asn Ala
485 490 495
Thr Asp Asp Pro Ala Ile Ser Asn Arg Thr Gln Gly Glu Ser Glu Lys
500 505 510
Lys Asn Asn Gln Asp Leu Lys Pro Ala Gln Asn Asp Leu Asp Phe Val
515 520 525
Arg Ala Asp Val
530
<210>32
<211>532
<212>PRT
<213>Nipah virus
<400>32
Met Ser Asp Ile Phe Glu Glu Ala Ala Ser Phe Arg Ser Tyr Gln Ser
1 5 10 15
Lys Leu Gly Arg Asp Gly Arg Ala Ser Ala Ala Thr Ala Thr Leu Thr
20 25 30
Thr Lys Ile Arg Ile Phe Val Pro Ala Thr Asn Ser Pro Glu Leu Arg
35 40 45
Trp Glu Leu Thr Leu Phe Ala Leu Asp Val Ile Arg Ser Pro Ser Ala
50 55 60
Ala Glu Ser Met Lys Val Gly Ala Ala Phe Thr Leu Ile Ser Met Tyr
65 70 75 80
Ser Glu Arg Pro Gly Ala Leu Ile Arg Ser Leu Leu Asn Asp Pro Asp
85 90 95
Ile Glu Ala Val Ile Ile Asp Val Gly Ser Met Val Asn Gly Ile Pro
100 105 110
Val Met Glu Arg Arg Gly Asp Lys Ala Gln Glu Glu Met Glu Gly Leu
115 120 125
Met Arg Ile Leu Lys Thr Ala Arg Asp Ser Ser Lys Gly Lys Thr Pro
130 135 140
Phe Val Asp Ser Arg Ala Tyr Gly Leu Arg Ile Thr Asp Met Ser Thr
145 150 155 160
Leu Val Ser Ala Val Ile Thr Ile Glu Ala Gln Ile Trp Ile Leu Ile
165 170 175
Ala Lys Ala Val Thr Ala Pro Asp Thr Ala Glu Glu Ser Glu Thr Arg
180 185 190
Arg Trp Ala Lys Tyr Val Gln Gln Lys Arg Val Asn Pro Phe Phe Ala
195 200 205
Leu Thr Gln Gln Trp Leu Thr Glu Met Arg Asn Leu Leu Ser Gln Ser
210 215 220
Leu Ser Val Arg Lys Phe Met Val Glu Ile Leu Ile Glu Val Lys Lys
225 230 235 240
Gly Gly Ser Ala Lys Gly Arg Ala Val Glu Ile Ile Ser Asp Ile Gly
245 250 255
Asn Tyr Val Glu Glu Thr Gly Met Ala Gly Phe Phe Ala Thr Ile Arg
260 265 270
Phe Gly Leu Glu Thr Arg Tyr Pro Ala Leu Ala Leu Asn Glu Phe Gln
275 280 285
Ser Asp Leu Asn Thr Ile Lys Ser Leu Met Leu Leu Tyr Arg Glu Ile
290 295 300
Gly Pro Arg Ala Pro Tyr Met Val Leu Leu Glu Glu Ser Ile Gln Thr
305 310 315 320
Lys Phe Ala Pro Gly Gly Tyr Pro Leu Leu Trp Ser Phe Ala Met Gly
325 330 335
Val Ala Thr Thr Ile Asp Arg Ser Met Gly Ala Leu Asn Ile Asn Arg
340 345 350
Gly Tyr Leu Glu Pro Met Tyr Phe Arg Leu Gly Gln Lys Ser Ala Arg
355 360 365
His His Ala Gly Gly Ile Asp Gln Asn Met Ala Asn Arg Leu Gly Leu
370 375 380
Ser Ser Asp Gln Val Ala Glu Leu Ala Ala Ala Val Gln Glu Thr Ser
385 390 395 400
Ala Gly Arg Gln Glu Ser Asn Val Gln Ala Arg Glu Ala Lys Phe Ala
405 410 415
Ala Gly Gly Val Leu Ile Gly Gly Ser Asp Gln Asp Ile Asp Glu Gly
420 425 430
Glu Glu Pro Ile Glu Gln Ser Gly Arg Gln Ser Val Thr Phe Lys Arg
435 440 445
Glu Met Ser Ile Ser Ser Leu Ala Asn Ser Val Pro Ser Ser Ser Val
450 455 460
Ser Thr Ser Gly Gly Thr Arg Leu Thr Asn Ser Leu Leu Asn Leu Arg
465 470 475 480
Ser Arg Leu Ala Ala Lys Ala Ala Lys Glu Ala Ala Ser Ser Asn Ala
485 490 495
Thr Asp Asp Pro Ala Ile Ser Asn Arg Thr Gln Gly Glu Ser Glu Lys
500 505 510
Lys Asn Asn Gln Asp Leu Lys Pro Ala Gln Asn Asp Leu Asp Phe Val
515 520 525
Arg Ala Asp Val
530
<210>33
<211>18234
<212>DNA
<213>Hendra virus
<400>33
accgaacaag gggaaatatg gatacgtgtt aaaaaactgc gtatgtttaa aacttaggaa 60
ccaagacagt gacaattggt cttggtattg gacaattgtt caaggttcca aaatgagtga 120
tatatttgac gaggcggcta gtttcagaag ctatcaatcg aaactcggtc gagatgggcg 180
ggcaagtgcg gcaacagcta ctttgactac taagataaga atttttgtac cagcaactaa 240
tagcccagaa ctgagatggg agttgacttt gttcgctctc gatgtaatca gatcaccaag 300
tgcagcagaa tcaatgaaga ttggtgctgc tttcactctg atatcaatgt actcagaaag 360
acctggcgct ttaatcagga gcctccttaa cgacccagac atagaagctg tcatcataga 420
tgtgggttcc atgctcaacg ggatccctgt gatggaaagg agaggggaca aagcacaaga 480
ggagatggaa ggtttgatga ggattctgaa gactgcacgt gaaagcagca agggaaagac 540
cccgtttgta gatagcagag cctatggact gaggatcacc gatatgagca ctcttgtgtc 600
agccgtcatt accatcgaag cccaaatttg gatcctgatt gcaaaggcag tgactgctcc 660
agatacagcc gaggaaagtg agaccagaag atgggcaaag tatgttcaac aaaagagggt 720
caatccattc tttgccttaa cccagcaatg gctgacagag atgaggaatc tcctctcaca 780
aagtctctca gtcagaaaat tcatggtgga aattctgatg gaggtaaaga aaggcggatc 840
agctaaagga agggccgttg agataatatc tgacatagga aattatgtcg aggaaacagg 900
aatggccggc ttctttgcga ctatcagatt cggtcttgaa acaagatacc ctgcacttgc 960
cctcaatgag ttccagagtg atctcaatac catcaaaggg ctgatgctgc tctacagaga 1020
aatagggcct cgagcgccat acatggtgct ccttgaggaa tctattcaga caaagtttgc 1080
accaggtggt tatccactcc tgtggagctt tgctatgggt gttgcaacca ctatcgaccg 1140
atccatgggt gctctcaaca tcaatcgtgg ttatcttgaa cctatgtatt tcagacttgg 1200
acagaaatca gcaagacatc atgctggcgg gattgatcaa aacatggcta ataaacttgg 1260
attaaattct gatcaagttg cagaactggc tgctgcagtt caagaaacat cagttgggag 1320
acaggacaac aacatgcagg caagagaagc caaatttgca gctgggggag ttcttgtggg 1380
gggtggcgaa caagatattg atgaggaaga agagcctatc gaacatagtg gtaggcagtc 1440
agtgactttc aagagagaaa tgagcatgtc atctctcgca gacagtgtcc caagcagttc 1500
agtgagtaca tccggaggaa cgagattgac aaattctcta ttgaatctca gatccagatt 1560
agctgcaaag gctatcaaag agagtactgc acagagctca tcggaaagaa acccacctaa 1620
taaccgccct caggcagact caggaaggaa agatgaccag gaacccaaac ctgcccaaaa 1680
tgatttggac tttgttagag cagacgtgtg acgcttaatc agaaactcat ataagctcca 1740
aatcattctc tatgacacac tcttataaat aaatttaaga gagtggattg agattgagtt 1800
atggacctgt gtcataatat acctttgtct tttaataaat tacttatacc acccacatat 1860
gatgaatatg agttcgcagt agtttgtctt ataatttaaa tcactctaaa gccatgcttg 1920
aacctcaact ttggatagtc acactgcaat gaatacattt agatatcctc agagtattta 1980
actcactaaa tctcagtagt agtaagtctc actggtgatg ggatacaagt gcaagttcct 2040
tttcaaatgg tgctcaaatc taaggatcct cttgctgaat aggctatcta ctatgcactg 2100
gaaaatataa tgaactttac ctcatacata actaaatgat caaagtttgt actgagtaca 2160
tctattaaag tttataacag gctaggggca acacctagaa ataatgtaag tctgttattt 2220
tagataacag tatatgagtg gcatgggtct agacaacttg atcattatat taagaaaaac 2280
ttaggatcca agaccataaa tctaggatcc tttacaatct cagaccctgg tgcaaggtta 2340
tatatagagc ccaattctcc taattaaaca gtgtctcagg ttgacaaatg gacaagttgg 2400
atctagtcaa tgatggcctc gatattattg actttattca gaagaaccaa aaagaaatac 2460
aaaagacata cggacgatca agcatccaac aaccaagtac caaagacagg acaagagcat 2520
gggaggactt cttgcagagc accagtggag aacatgaaca ggctgagggg ggaatgccta 2580
agaatgatgg aggtactgaa ggaagaaatg tggaggatct atccagtgtt acttcctcag 2640
atggaactat tggacaaaga gtgtcaaaca cccgagcttg ggcagaagac ccagatgaca 2700
tacaactgga cccaatggtt acagacgttg tataccatga tcatggagga gaatgtaccg 2760
gacatggacc ttcttcaagc cctgagagag ggtggagtta tcacatgtca ggaacacacg 2820
atgggaatgt acgtgctgta cctgatacaa aggtgttgcc caatgctccc aaaactacag 2880
ttcctgaaga agttagggaa attgatttaa ttgggttgga ggacaaattt gcatcagcgg 2940
gattaaatcc agctgcagtt ccctttgtcc ccaagaatca atcaacccca actgaggagc 3000
ctccagtcat cccggagtat tactatggat ctgggaggag aggagatctc tcaaaatctc 3060
ctcctagagg taatgtcaat ctggacagca tcaagattta cacttcggac gacgaggatg 3120
aaaatcagct ggagtatgag gatgagtttg ccaaaagctc aagtgaggtt gtcattgaca 3180
ctactcctga ggacaatgat tctatcaacc aggaggaagt agttggggac ccgtctgatc 3240
agggtttaga gcatcctttc cctttgggga aattcccgga gaaagaagaa actcctgatg 3300
tacgcagaaa ggattcccta atgcaggaca gttgtaagag aggaggagta ccgaaaagac 3360
tgcccatgtt atctgaagag ttcgagtgct ccggatcaga tgatccaata atacaagaat 3420
tagaacgaga agggtctcat ccaggaggtt cattacgttt gagggaacca cctcagtcgt 3480
cagggaattc cagaaatcaa cctgaccgtc agctgaagac aggtgatgca gcatcacccg 3540
gaggcgtcca gagaccaggc acaccgatgc ctaaatcaag gatcatgccc attaaaaagg 3600
gcacagacgc gaagtctcaa tatgttggga cggaagacgt gcctgggtcg aagagtggtg 3660
caacccggta tgttcgcgga ttacccccca accaagaaag caagagtgtt actgcggaga 3720
atgtccaact gagtgctccc agtgctgtca cgaggaatga aggacacgac caggaagtta 3780
ccagcaacga agatagtctg gatgacaaat atattatgcc atcagatgat tttgctaaca 3840
ctttcttacc ccatgacact gataggttaa attatcatgc agaccatctc aatgattacg 3900
atttagagac cttgtgtgag gagtcagtat tgatgggcat cgttaacgca atcaaactca 3960
tcaacattga tatgaggttg aatcacattg aggaacagat gaaagagata cccaagatca 4020
tcaacaaaat agactccatt gatcgagtgt tggctaagac caatactgca ttgtcgacaa 4080
tagaaggaca cctagtctcg atgatgatca tgataccagg gaaaggcaag ggtgaaagga 4140
aagggaaaac aaatccggag ctaaaacctg taattgggag aaatatccta gaacagcaag 4200
agttattttc attcgacaat ctcaagaatt tcagagatgg gtcattgact gatgagccct 4260
atggaggggt ggcccgaata agagatgatt tgatcctgcc tgaactcaac ttcagtgaga 4320
caaatgcatc acaatttgtt cctttggcag atgatgcatc taaggatgtc gtgagaacta 4380
tgattaggac tcacatcaag gacagagagt tgaggtctga gttgatggat tatctaaatc 4440
gagcagaaac agatgaagag gttcaggagg tggccaatac agtcaatgat attattgatg 4500
ggaacatcta aacatgtcag taatttgata gagtagcagt cagaccaaga tcatatatta 4560
gaggtacaca tcataatggc cagtgtctta atctcaacca ttcataaacc gtcagtgttt 4620
cccttataaa ctccattatg tagtttatta atatctagag ctattcttta ttaagtaata 4680
aatctgtatt agattattat ataatctcta taatcaacac gcctattact tcatgcactt 4740
actatcacca tttaataata gttcagatca cccctcaaaa tctgggttgt gccaagtcat 4800
tttcctcacc atataatcca cgcagtatgc attcaaatac taaaccacat tcagacagag 4860
ttagacattc aagcaatcaa taactatatt aactataaga atgatgaatc agccctgcgc 4920
caacaaggca atgtcaatca cttgaatgca ggggtagacc ccagaatcca ttaagaaaaa 4980
cttaggagac aggtataata ttctcatacc tgtccattca ccttggactt aaagagagac 5040
aatcattcag ccccgtccaa gacacacctg agtggccaga accatggatt ttagtgtgag 5100
tgataacctt gatgatccaa tagaaggtgt ttcagatttt agtcccacct catgggagaa 5160
tggagggtat ctagataaag tggagccaga aattgataag catggcagta tgatacctaa 5220
gtacaagatc tataccccag gtgcaaatga gaggaaattc aacaactaca tgtacatgat 5280
ttgctatggc ttcgtcgaag acgttgagag atcaccagaa tctgggaagc gcaagaaaat 5340
caggaccatt gccgcgtacc ctcttggtgt tggcaagagc acttctcacc ctcaggatct 5400
tttagaagag ctatgttctt tgaaagtcac tgtcagaaga acagctgggg ctacagagaa 5460
aattgtgttc gggtcctctg gcccgcttca tcatctctta ccatggaaga agattctgac 5520
tggcggatca atattcaatg ctgtcaaggt ttgccgcaat gtggatcaga ttcaactgga 5580
gaatcaacaa tcattgagga tttttttctt gagtattact aaattgaatg actccggcat 5640
ctacatgatc ccaaggacaa tgctagaatt cagaaggaac aacgctattg ctttcaatct 5700
tctagtatac ctcaaaatcg atgcagatct tgcaaaagct ggaatccaag gaagcttcga 5760
caaagacgga accaaagtgg catctttcat gctccatctc gggaattttg tccgacgagc 5820
tgggaagtac tattctgtcg aatactgcaa gagaaagatt gacagaatga agctccagtt 5880
ctctcttggt tcaattgggg gtctaagctt acacatcaaa attaatggag tgatcagtaa 5940
gagattattt gcccagatgg gtttccagaa gaacctttgc ttttctctaa tggacatcaa 6000
tccttggctc aacagactga cttggaacaa cagttgtgag atcagcaggg tggcagctgt 6060
actgcagccc tcggtgccac gtgaattcat gatctatgac gatgtgttca ttgacaacac 6120
agggaagatc ctaaaggggt gaaataaaac attcacttgg ttattgtaga ttgataaatt 6180
aaccagataa ccactttaat cttcattcta aagaatgcca cagtgtcagt aagttacaca 6240
ggtacaatac ataatctaaa ttcatgtctt cccttcaaag gattgatttg gacctttaaa 6300
atagtagagg taagaattga gtatatccaa tattaagaaa aacttaggag ccaagttttg 6360
gtctctttag attaatcaag ctgtttgtca atctgtattc gtaaggttgt taaaataatc 6420
tgtgtctatc ctttagatat ccacttgggt tgtagatata gtcaatcaac tggtctgaca 6480
agtcataccc caaacaatcc aaatctggaa tcacctgttc gctcagtagt aatatcttgg 6540
caaacactac actaacctcg tctgtactgt ctgaaagtgg tcaggattga ttgacagtta 6600
ttatttatct ttaagcaatg gctacacaag aggtcaggct aaagtgtttg ctctgtggga 6660
tcatagttct ggttttgtca ttagaagggc tagggatact acattatgag aaacttagta 6720
agatagggct ggttaaaggt attacaagaa agtacaagat taagagtaac cctttgacca 6780
aggatattgt gatcaaaatg atccctaatg tctcgaatgt ctcaaagtgc accgggactg 6840
ttatggagaa ttacaaaagc agactcacag ggattctctc accaatcaaa ggcgccatcg 6900
aactgtacaa taataacacg catgacctag ttggtgatgt caagcttgca ggtgtggtga 6960
tggcagggat tgcaatcggg atagctactg ctgcacaaat cacagcaggt gttgccttat 7020
atgaggcaat gaagaacgca gacaatatca ataaactcaa gagcagcata gagtctacaa 7080
atgaggctgt tgtcaaatta caggaaacag ctgagaaaac agtctacgtc cttactgctc 7140
ttcaagatta catcaacact aaccttgttc ctacaataga tcaaattagc tgcaagcaaa 7200
cagagctcgc attagacttg gcgttgtcta agtatctgtc tgatctgctc tttgttttcg 7260
gacctaactt acaggatcca gtctctaatt ccatgactat ccaagcaata tctcaagcat 7320
ttgggggcaa ttacgaaacc ttactgagaa cgcttggtta cgcgaccgag gacttcgacg 7380
accttttaga aagtgatagc atagcaggcc agatagtcta tgtagatctc agtagctatt 7440
acataatagt aagggtgtat tttcccatac taacagagat ccaacaggct tatgtgcagg 7500
agttgcttcc agtgagtttt aataacgata attcagaatg gatcagcatt gtcccgaatt 7560
tcgtgctgat taggaacacg ctgatttcaa atatagaagt caagtactgc ttaatcacca 7620
agaaaagtgt gatttgtaat caggactatg ctacacccat gacggctagc gtgagagaat 7680
gcttgacagg atccacagat aagtgcccaa gggagttagt agtctcatcc catgttccaa 7740
gatttgccct ctcaggagga gtcttgtttg caaattgtat aagtgtgaca tgtcagtgtc 7800
agactactgg gagggcaata tctcaatcag gggaacagac actactgatg attgacaata 7860
ctacctgcac aacagttgtt ctaggaaaca taatcataag ccttggaaaa tatttgggat 7920
caataaatta caattctgag agcattgctg ttgggccacc agtctataca gacaaagttg 7980
atatctcaag tcagatatct agtatgaatc aatcactaca acaatctaag gattacatta 8040
aagaagctca aaagatcttg gacactgtga atccgtcgtt gataagtatg ctatcaatga 8100
tcatccttta tgttttgtcc attgcagcac tgtgcattgg tctgatcact ttcataagct 8160
ttgtaatagt tgagaaaaag agagggaatt acagcaggct agatgatagg caagtgcgac 8220
cggtcagtaa tggtgatctg tattatattg gaacataaaa tacactacaa tcaatccaat 8280
tgttttatct ttactaatga tataaacgat acttaatcat cagctaccta agtgctcata 8340
tagctctcat gtatattctt ttattattgt taacaacagg tctactaaat agtttaatcc 8400
aataaattga ggataatcaa ttcttgcatt gcataaactc gttaggttat atataacaaa 8460
aattattttt ttctttttga aagattgtat agtctggtca ttctattata attgatataa 8520
tgattctttg atcatattga tcaatgtacc tatttgaatc aatcgacatt ggattctcta 8580
ttcttatgta atcccagatg gatcaaggct agtcatacct taaatcacaa gctgtattat 8640
tacttactgt gatggataaa gggactttac aaaaaactta ggacccaagt ccttaaccac 8700
attctaatgt gagggagaat taaatgtaca ttgagactga caatctaata tacagagtgt 8760
ttgatcaagt taaaacatca attgtcagga attcattgat aaaccaactt gttagttaga 8820
attaagaaaa tataagagct aatactcgca agtcatttgc ttcttcaaga gcctgtctca 8880
actatcaagt gaatacatga tctaaaacta gtatgatggc tgattccaaa ttggtaagcc 8940
tgaacaataa tctatctggt aaaatcaagg atcaaggtaa agttatcaag aattattacg 9000
gcacaatgga catcaagaaa attaacgatg ggttattaga tagtaagata cttggggcgt 9060
ttaacacagt gatagctttg ttgggatcaa tcatcatcat tgtgatgaat atcatgataa 9120
ttcaaaatta caccagaacg actgataatc aggcactaat caaagagtca ctccagagtg 9180
tacagcaaca aatcaaagct ttaacagaca aaatcgggac agagataggc cccaaagtct 9240
cactaattga cacatccagc accatcacaa ttcctgctaa catagggtta ctgggatcca 9300
agataagtca gtctaccagc agtattaatg agaatgttaa cgataaatgc aaatttactc 9360
ttcctccttt aaagattcat gagtgtaata tctcttgtcc gaatcctttg cctttcagag 9420
aataccgacc aatctcacaa ggggtgagtg atcttgtagg actgccgaac cagatctgtc 9480
tacagaagac aacatcaaca atcttaaagc ccaggctgat atcctatact ctaccaatta 9540
ataccagaga aggggtttgc atcactgacc cacttttggc tgttgataat ggcttcttcg 9600
cctatagcca tcttgaaaag atcggatcat gtactagagg aattgcaaaa caaaggataa 9660
taggggtggg tgaggtattg gataggggtg ataaggtgcc atcaatgttt atgaccaatg 9720
tttggacacc acccaatcca agcaccatcc atcattgcag ctcaacttac catgaagatt 9780
tttattacac attgtgcgca gtgtcccatg tgggagatcc tatccttaac agtacttcct 9840
ggacagagtc actgtctctg attcgtcttg ctgtaagacc aaaaagtgat agtggagact 9900
acaatcagaa atacatcgct ataactaaag ttgaaagagg gaagtacgat aaggtgatgc 9960
cttacggtcc atcaggtatc aagcaagggg atacattgta ctttccggcc gtcggttttt 10020
tgccaaggac cgaatttcaa tataatgact ctaattgtcc cataattcat tgcaagtaca 10080
gcaaagcaga aaactgtagg ctttcaatgg gtgtcaactc caaaagtcat tatattttga 10140
gatcaggact attgaagtat aatctatctc ttggaggaga catcatactc caatttatcg 10200
agattgctga caatagattg accatcggtt ctcctagtaa gatatacaat tccctaggtc 10260
aacccgtttt ctaccaggca tcatattctt gggatacgat gattaaatta ggcgatgttg 10320
ataccgttga ccctctaaga gtacagtgga gaaataacag tgtgatttct agacctggac 10380
agtcacagtg tcctcgattt aatgtctgtc ccgaggtatg ctgggaaggg acatataatg 10440
atgcttttct aatagaccgg ctaaactggg ttagtgctgg tgtttattta aacagtaacc 10500
aaactgcaga gaaccctgtg tttgccgtat tcaaggataa cgagatcctt taccaagttc 10560
cactggctga agatgacaca aatgcacaaa aaaccatcac agattgcttc ttgctggaga 10620
atgtcatatg gtgtatatca ctagtagaaa tatacgatac aggagacagt gtgataaggc 10680
caaaactatt tgcagtcaag atacctgccc aatgttcaga gagttgattg accaaaagag 10740
caataaatat tatattataa ttatatcagt caaattgtaa acatccttct tataatatac 10800
tcaaaaaatt aaaaattccc ccaagaaaat actaaatgta atcaaactca ataaaccctg 10860
aaatattgat agtttacaga gatcatatga ctctttgtat aattctttat agaactccta 10920
aatatcaaag agcgataaac ttgctaccct gtacacttaa agcagatact gtgattaact 10980
ctgtaagttt atcagataat cctattagtg acatactgag ggttattctt gtataattct 11040
tctgaagatg tgattacaga taatctttca atctagtcaa gtaagcctga tttcctaatt 11100
ataatctccc tcttaggaga aggagaacgt atcaatgttc actatatact aagtctttta 11160
ttttaaattt ttcttattgt cttcagatga gccttattac ttcatctcat cagattataa 11220
attttctaat aaattaagaa aaacttagga cccaagtcct ttaaacgtgc atgattgaga 11280
tagtcagaaa ttggagctaa tttgaatact agtgagtaaa ttaggagtag tagtattgaa 11340
ccacctcata tcttatttct tctgaactgc attaaaattt gtacccagga tataacaaca 11400
tggctcatga attatctatc tctgatatca tctatcctga atgtcacctt gacagtccta 11460
tagtatcagg gaaattgatt tcagctatag aatatgcgca gttgagacat aaccaaccga 11520
atggggacaa gagattaact gaaaatatca aaattaactt acagggcaaa agaagaagtg 11580
tgtatatatc cagacaatct agacttggaa attacattag agacaatata aagaatttga 11640
aggaatttct acatgtatca tatcccgagt gcaataaatc tttattctct ctcaagtctc 11700
ctggaatgac aagtaagttg agtaatatca tgaagaaatc atttaaagca tacaatatcg 11760
tgagcagaaa aatcatagag atgttacaaa acatcaccag gaatctcata actcaggacc 11820
aaaaggatga agttttaggt atttatgagc aggacagact cagtaacata gggaaataca 11880
tgagtcaatc gcaatggtat gagtgttttc tcttctggtt cacaatcaag acagagatga 11940
gagctgttat caagaactca cagaaaccta agtttagatc tgactcttgt attattcata 12000
tgaaagataa taacatggaa attgtaatga acccaaatct tgtctgcatt tacaaaaatg 12060
ataaggatgg aaagaggtgt tactacctta caccagaaat tgttctaatg tgttgtgatg 12120
tcttggaagg ccggatgatg atcgagactt cgataaaatc tgatatcaaa taccaatctt 12180
taattaccag atccaacgct ttgtggacat tcattgattc actcttcccg attatgggta 12240
ataggatata caatatagta tctatgatag agcctttagt cttggcactt ttacagttga 12300
aagatgaggc gcgcatccta agaggggcat ttctacatca ttgcattaaa gagatccacc 12360
aagagttgat tggatgtggg tttactgatc aaaaaacaag atctatcttt atcgatgatc 12420
ttctatcagt aatgaatatt gataatatcc atctgttagc tgagttcttt tctttcttcc 12480
gaacctttgg tcatccaatt ctcgaagcca aaaccgcagc agacaaagtt agagagcata 12540
tgctagcaga taaggtcctt gaatacgggc caatcatgaa agcacacgca gtcttttgtg 12600
gtacaattat aaatgggtac cgtgacaggc atcggggagc ttggcctcca ctttatctgc 12660
catcgcatgc atcgaaacac ataataagac tcaagaattc tggtgaatca ttaacagttg 12720
atgattgtgt taagaactgg gagtcatttt gtgggattca atttgattgc tttatggagt 12780
taaaattaga cagtgatttg agcatgtata tgaaagataa agcactatcc ccaataaagg 12840
aggaatggga tagtgtctat ccacgtgaag tacttaatta cacgccaccc agatctaccg 12900
aaccaagaag attggtggat gtatttgtca atgatgagaa tttcgatcct tataatatgt 12960
tagagtatgt ccttacagga gattatttaa ctgatgaaca attcaatgtt tcttatagtc 13020
taaaggagaa agaaactaaa caagcaggaa gattatttgc taaaatgaca tataagatgc 13080
gtgcctgtca ggtaattgct gaagctttga ttgcatcggg agtgggcaag tattttaaag 13140
agaacgggat ggttaaagat gaacatgagc tactcaaaac tttatttcaa ctgtcaatct 13200
cttcagtacc tcgaggcaat agtcagggca gagattccga attttccaat aacacagaga 13260
aaagtctcat atcactcaag agaacaacag gtaggcttct gaataatgaa gttccttgtc 13320
gaatgaacat tatgtcggct ttaattgaca aaaatcagtc agaccaaaag aagcataata 13380
ttctcccaaa cacacgaaat cgccacaagt gtgataacac tagccaaaca ttccttgact 13440
atcatatgga attcagtccc tacaagtcag acagaatgga taggacagag acatctgatt 13500
tctcgaaata tgatgatgga acaggaacaa aatttgacac ggtgagtgct ttcctaacta 13560
ctgacttgaa gaaattctgt ctcaattggc gatatgaatc catggctatt tttgcagaaa 13620
gacttgacga aatttatggc ttacccggtt tctttaactg gatgcacaaa cgattagaga 13680
agtcagtcat ttatgtcgca gatcctaatt gcccaccaga catcgggaaa catataaacc 13740
ttgatgatac tcctgaagat gacatattta tccattcacc aaaaggcggg attgaaggtt 13800
atagtcagaa gacttggacg atcgcaacaa ttccctttct gtttctcagt gcctacgaaa 13860
caaatacaag gattgcagct atagttcagg gagacaatga gtcaattgcc ataactcaga 13920
aggtccaccc taatttacct tacaaagtca agaaagaaat ttgtgcaagg caagctcaat 13980
tatattttga caggctcaga atgaatctta gggccttagg actcaactta aaagcaacag 14040
agaccattat aagtacacac ttatttgtct actctaagaa aatacattat gatggtgcag 14100
tattatcaca agccctcaaa tcaatgtcta ggtgttgttt ttggtcagag acattggtgg 14160
acgagacaag atctgcttgc agcaatatta gtactaccat agctaaggcg attgagaatg 14220
gattatcaag aaatgtgggt tactgcatca acgtcttgaa ggtcatacag caactgctta 14280
tttctactga gtttagcatc aacgagacat tgactgctga tgtaacatct ccaatatcca 14340
ataatctcga ttggctggta actgcatccc gaattcccgc accaattgga ggtttcaatt 14400
acttgaattt atcaagaata tttgtaagaa atataggaga ccctgtgact gcatcactag 14460
ccgatctaaa gagaatgata gaacatgact tgatgacaga taaagttctc caaaaagtga 14520
tgaatcaaga accaggcgat gcaagcttcc tagattgggc tagtgatccg tattcaggaa 14580
atctaccgga ttcacagagt attacaaaaa cgatcaagaa tataacggct aggactatat 14640
tgaggacatc accgaatcca atgctgaagg gtctattcca tgataaatca ttcgaggaag 14700
atctggagct tgctactttc ctaatggatc gtagaattat attgcccaga gccgcacatg 14760
aaattttaga taattctttg acaggtgcta gagaggaaat tgctggatta cttgacacaa 14820
caaaggggct aataagatca ggattgaaaa agagcgggat acaacctaag ttagtgtcta 14880
gactgtctaa ccatgattat aatcagttcc tgatactcaa tagacttttg tccaataaaa 14940
agagaaacga tcttatatcc cctaaaacat gttcagtgga tttggccaag gctcttcgtt 15000
gtcatatgtg gagagatctt gctcttggtc gatcaatcta tggacttgaa gtgcctgacg 15060
ctttggaagc aatgactgga agatatatta cagggagtat ggaatgccaa ctttgtgatc 15120
aagggaatac tatgtacggt tggtttttcg tgccacgaga ttcacaatta gaccaagtga 15180
ataaagagca ctcatccatt agagtacctt acgtcggctc aagcactgat gagaggtcag 15240
atatcaaact aggaaacgtc aagagaccga caagagcact gagatctgcc atcgaattgc 15300
aactgtttat acctgggcat acggagattc tgaagaaagt tggtatgagg cttggtactt 15360
ggcttcccag aggggtcaac attgatatag atgtgttaaa agctataact cctgtctcaa 15420
cttccaacaa tctatctcac agactcagag atagatctac acagttcaag ttgccaggta 15480
gtgtccttaa tagggtttct agatatgtca acataagtaa tgataattta gacttcagag 15540
tggaaggtga aaaggtcgac accaatctaa tataccaaca aacaatgcta ctcggcttgt 15600
cagtattgga aggtaaattt cggttgagaa cagaaacaga tgattataac ggaatatatc 15660
atctgcatgt cagagataat tgttgtgtca aggaggttgc tgatattggt ggtgttaatg 15720
ctgagctacc tgtcccagaa tacactgaag tagaaaataa taggctcata tatgacccag 15780
atcctgtttc tgaaattgat tgtgatcgct tatcaaagca agagtctaag gcgagagaat 15840
tagactttcc tctttggtcc accgaagaac tccatgatgt gttagctaag actgtggctc 15900
aaacagttct cgagatcata actaaagccg ataaagatgt cctcaagcaa cacttagcta 15960
tagactcaga tgacagtatc aatagtctta taacagagtt tttaatggtg gatcctgagc 16020
tttttgccct ttatttaggt caatcaattt cagtcaagtg ggcatttgaa atccatcata 16080
gacgcccaca tggacgacat acaatggtag atttgttgtc tgatctcata tccaatacat 16140
cgaaacatac atacaaagtc ttgtctaatg cactgtcaca tcccagagtg ttcaagaggt 16200
ttgtgaattg cggcttgctt cttcccactc agggtcctta tcttcaccaa caagattttg 16260
agaaattatc tcagaacctc ttaatcacat cttatatgaa ttatcttatg aattggtgtg 16320
acttcaaaaa attccctttt ttgatagctg aacaagatga agcagtcgta gaattaaggg 16380
aagatataat aacatcaaaa cacctgtgta tgataattga tctgtacgca aatcaccata 16440
agccaccttg gataattgat ctcaacccac aagagaagat atgtgttctt agggacttta 16500
tatctaagtg ccgtcatact gatgtgtcat ctaggtcttg gaatatcact gatctggatt 16560
ttatggtttt ttatgcttcg ttgacttatc taagaagagg aataatcaag cagctcagaa 16620
ttaggcaagt gacagaagta atcgacacca ctacaatgct aagagataat attttagtag 16680
aaaacccgcc tattaaaact ggagtactag atattagagg ttgcataata tataatctag 16740
aagaaatctt atcaatgaat actaagtcca cgtcaagaaa ggtgttcaat ttgggttcaa 16800
agttgtcagt tgaaaaccac aaatatagaa ggataggatt aaattcgtct tcttgctaca 16860
aggcattaaa tctgtcaccg ttgattcaac gctacctgcc cgcaggttca caacgattat 16920
ttgtaggtga agggtcagga agtatgatgc tattatatca acagacttta ggatgttcga 16980
tatcctttta caactccggt attgatggtg attatatacc agggcagaga gaattgagat 17040
tattcccttc cgaatattca atagctgaag acgatccgtc acaatctgat aagctaaaag 17100
gtctagtggt cccattattt aatggaaggc ccgaaaccac gtggataggg aacttagatt 17160
catacgagta cataatcaac agaacagccg gtcgtaacat cggattggta cactcagata 17220
tggaatcagg catagacaaa caagtggaag aaattatgat agagcattca caccttatct 17280
caatagcaat caatgtaatg atagaagacg gtgttcttgt atctaaaata gcatttgcac 17340
cagggttccc catatccagg ctactgaata tgtatcgttc ctattttggg ttagtattgg 17400
tgtgtttccc agtgtatagt aatccagagt ccacagaagt gtatctcatc tgcttgcaga 17460
aaacaattaa gaccataatt ccacctcaaa aagttcttga tcattcatat ctcagtgatg 17520
aaatcaatga tcaaggtatt acatctgtta ttttcaagat caaaaacatt caatcaaaac 17580
aattccatga agatctagtg aaacactatc aggtcgaaca accgtttttt gttcctagtc 17640
atatcacatg tgatgaaaag ttgcttatgc aggctggttt gaaaatgaac ggaccagaaa 17700
tcttgaagaa tgaagtggga tatgatatag gatcagatat caatacactc aggagcacta 17760
ttattatctt acttaatgaa gctatgaact attttgacga tgagaggagt ccttctcatc 17820
atctagagcc ctttccagtc ctagagaaaa cgagagtcaa aactatcatg gggagagtca 17880
cgagaaaagt aactgtttat tctcttataa agttgaagga aactaaaagc ccggaacttt 17940
ataacatcaa aaattacata aggagaaaag ttttgattct agacttcaga tcccatacta 18000
tgattaaact gctacctaaa ggaatgaaag agagaagaga gaaaagcgga ttcaaagaaa 18060
tttggatatt tgacttatct aatagggaag tcaaaatctg gtggaagatc ataggttacc 18120
tctctttagt ctgaatcgaa ctaccacacc catcaccaat gaagcccgaa caagtctaca 18180
tagatcagtt attaagaaaa acttaataac gatctctctt tacccttgtt cggt 18234
<210>34
<211>532
<212>PRT
<213>Hendra virus
<400>34
Met Ser Asp Ile Phe Asp Glu Ala Ala Ser Phe Arg Ser Tyr Gln Ser
1 5 10 15
Lys Leu Gly Arg Asp Gly Arg Ala Ser Ala Ala Thr Ala Thr Leu Thr
20 25 30
Thr Lys Ile Arg Ile Phe Val Pro Ala Thr Asn Ser Pro Glu Leu Arg
35 40 45
Trp Glu Leu Thr Leu Phe Ala Leu Asp Val Ile Arg Ser Pro Ser Ala
50 55 60
Ala Glu Ser Met Lys Ile Gly Ala Ala Phe Thr Leu Ile Ser Met Tyr
65 70 75 80
Ser Glu Arg Pro Gly Ala Leu Ile Arg Ser Leu Leu Asn Asp Pro Asp
85 90 95
Ile Glu Ala Val Ile Ile Asp Val Gly Ser Met Leu Asn Gly Ile Pro
100 105 110
Val Met Glu Arg Arg Gly Asp Lys Ala Gln Glu Glu Met Glu Gly Leu
115 120 125
Met Arg Ile Leu Lys Thr Ala Arg Glu Ser Ser Lys Gly Lys Thr Pro
130 135 140
Phe Val Asp Ser Arg Ala Tyr Gly Leu Arg Ile Thr Asp Met Ser Thr
145 150 155 160
Leu Val Ser Ala Val Ile Thr Ile Glu Ala Gln Ile Trp Ile Leu Ile
165 170 175
Ala Lys Ala Val Thr Ala Pro Asp Thr Ala Glu Glu Ser Glu Thr Arg
180 185 190
Arg Trp Ala Lys Tyr Val Gln Gln Lys Arg Val Asn Pro Phe Phe Ala
195 200 205
Leu Thr Gln Gln Trp Leu Thr Glu Met Arg Asn Leu Leu Ser Gln Ser
210 215 220
Leu Ser Val Arg Lys Phe Met Val Glu Ile Leu Met Glu Val Lys Lys
225 230 235 240
Gly Gly Ser Ala Lys Gly Arg Ala Val Glu Ile Ile Ser Asp Ile Gly
245 250 255
Asn Tyr Val Glu Glu Thr Gly Met Ala Gly Phe Phe Ala Thr Ile Arg
260 265 270
Phe Gly Leu Glu Thr Arg Tyr Pro Ala Leu Ala Leu Asn Glu Phe Gln
275 280 285
Ser Asp Leu Asn Thr Ile Lys Gly Leu Met Leu Leu Tyr Arg Glu Ile
290 295 300
Gly Pro Arg Ala Pro Tyr Met Val Leu Leu Glu Glu Ser Ile Gln Thr
305 310 315 320
Lys Phe Ala Pro Gly Gly Tyr Pro Leu Leu Trp Ser Phe Ala Met Gly
325 330 335
Val Ala Thr Thr Ile Asp Arg Ser Met Gly Ala Leu Asn Ile Asn Arg
340 345 350
Gly Tyr Leu Glu Pro Met Tyr Phe Arg Leu Gly Gln Lys Ser Ala Arg
355 360 365
His His Ala Gly Gly Ile Asp Gln Asn Met Ala Asn Lys Leu Gly Leu
370 375 380
Asn Ser Asp Gln Val Ala Glu Leu Ala Ala Ala Val Gln Glu Thr Ser
385 390 395 400
Val Gly Arg Gln Asp Asn Asn Met Gln Ala Arg Glu Ala Lys Phe Ala
405 410 415
Ala Gly Gly Val Leu Val Gly Gly Gly Glu Gln Asp Ile Asp Glu Glu
420 425 430
Glu Glu Pro Ile Glu His Ser Gly Arg Gln Ser Val Thr Phe Lys Arg
435 440 445
Glu Met Ser Met Ser Ser Leu Ala Asp Ser Val Pro Ser Ser Ser Val
450 455 460
Ser Thr Ser Gly Gly Thr Arg Leu Thr Asn Ser Leu Leu Asn Leu Arg
465 470 475 480
Ser Arg Leu Ala Ala Lys Ala Ile Lys Glu Ser Thr Ala Gln Ser Ser
485 490 495
Ser Glu Arg Asn Pro Pro Asn Asn Arg Pro Gln Ala Asp Ser Gly Arg
500 505 510
Lys Asp Asp Gln Glu Pro Lys Pro Ala Gln Asn Asp Leu Asp Phe Val
515 520 525
Arg Ala Asp Val
530
<210>35
<211>707
<212>PRT
<213>Hendra virus
<400>35
Met Asp Lys Leu Asp Leu Val Asn Asp Gly Leu Asp Ile Ile Asp Phe
1 5 10 15
Ile Gln Lys Asn Gln Lys Glu Ile Gln Lys Thr Tyr Gly Arg Ser Ser
20 25 30
Ile Gln Gln Pro Ser Thr Lys Asp Arg Thr Arg Ala Trp Glu Asp Phe
35 40 45
Leu Gln Ser Thr Ser Gly Glu His Glu Gln Ala Glu Gly Gly Met Pro
50 55 60
Lys Asn Asp Gly Gly Thr Glu Gly Arg Asn Val Glu Asp Leu Ser Ser
65 70 75 80
Val Thr Ser Ser Asp Gly Thr Ile Gly Gln Arg Val Ser Asn Thr Arg
85 90 95
Ala Trp Ala Glu Asp Pro Asp Asp Ile Gln Leu Asp Pro Met Val Thr
100 105 110
Asp Val Val Tyr His Asp His Gly Gly Glu Cys Thr Gly His Gly Pro
115 120 125
Ser Ser Ser Pro Glu Arg Gly Trp Ser Tyr His Met Ser Gly Thr His
130 135 140
Asp Gly Asn Val Arg Ala Val Pro Asp Thr Lys Val Leu Pro Asn Ala
145 150 155 160
Pro Lys Thr Thr Val Pro Glu Glu Val Arg Glu Ile Asp Leu Ile Gly
165 170 175
Leu Glu Asp Lys Phe Ala Ser Ala Gly Leu Asn Pro Ala Ala Val Pro
180 185 190
Phe Val Pro Lys Asn Gln Ser Thr Pro Thr Glu Glu Pro Pro Val Ile
195 200 205
Pro Glu Tyr Tyr Tyr Gly Ser Gly Arg Arg Gly Asp Leu Ser Lys Ser
210 215 220
Pro Pro Arg Gly Asn Val Asn Leu Asp Ser Ile Lys Ile Tyr Thr Ser
225 230 235 240
Asp Asp Glu Asp Glu Asn Gln Leu Glu Tyr Glu Asp Glu Phe Ala Lys
245 250 255
Ser Ser Ser Glu Val Val Ile Asp Thr Thr Pro Glu Asp Asn Asp Ser
260 265 270
Ile Asn Gln Glu Glu Val Val Gly Asp Pro Ser Asp Gln Gly Leu Glu
275 280 285
His Pro Phe Pro Leu Gly Lys Phe Pro Glu Lys Glu Glu Thr Pro Asp
290 295 300
Val Arg Arg Lys Asp Ser Leu Met Gln Asp Ser Cys Lys Arg Gly Gly
305 310 315 320
Val Pro Lys Arg Leu Pro Met Leu Ser Glu Glu Phe Glu Cys Ser Gly
325 330 335
Ser Asp Asp Pro Ile Ile Gln Glu Leu Glu Arg Glu Gly Ser His Pro
340 345 350
Gly Gly Ser Leu Arg Leu Arg Glu Pro Pro Gln Ser Ser Gly Asn Ser
355 360 365
Arg Asn Gln Pro Asp Arg Gln Leu Lys Thr Gly Asp Ala Ala Ser Pro
370 375 380
Gly Gly Val Gln Arg Pro Gly Thr Pro Met Pro Lys Ser Arg Ile Met
385 390 395 400
Pro Ile Lys Lys Gly Thr Asp Ala Lys Ser Gln Tyr Val Gly Thr Glu
405 410 415
Asp Val Pro Gly Ser Lys Ser Gly Ala Thr Arg Tyr Val Arg Gly Leu
420 425 430
Pro Pro Asn Gln Glu Ser Lys Ser Val Thr Ala Glu Asn Val Gln Leu
435 440 445
Ser Ala Pro Ser Ala Val Thr Arg Asn Glu Gly His Asp Gln Glu Val
450 455 460
Thr Ser Asn Glu Asp Ser Leu Asp Asp Lys Tyr Ile Met Pro Ser Asp
465 470 475 480
Asp Phe Ala Asn Thr Phe Leu Pro His Asp Thr Asp Arg Leu Asn Tyr
485 490 495
His Ala Asp His Leu Asn Asp Tyr Asp Leu Glu Thr Leu Cys Glu Glu
500 505 510
Ser Val Leu Met Gly Ile Val Asn Ala Ile Lys Leu Ile Asn Ile Asp
515 520 525
Met Arg Leu Asn His Ile Glu Glu Gln Met Lys Glu Ile Pro Lys Ile
530 535 540
Ile Asn Lys Ile Asp Ser Ile Asp Arg Val Leu Ala Lys Thr Asn Thr
545 550 555 560
Ala Leu Ser Thr Ile Glu Gly His Leu Val Ser Met Met Ile Met Ile
565 570 575
Pro Gly Lys Gly Lys Gly Glu Arg Lys Gly Lys Thr Asn Pro Glu Leu
580 585 590
Lys Pro Val Ile Gly Arg Asn Ile Leu Glu Gln Gln Glu Leu Phe Ser
595 600 605
Phe Asp Asn Leu Lys Asn Phe Arg Asp Gly Ser Leu Thr Asp Glu Pro
610 615 620
Tyr Gly Gly Val Ala Arg Ile Arg Asp Asp Leu Ile Leu Pro Glu Leu
625 630 635 640
Asn Phe Ser Glu Thr Asn Ala Ser Gln Phe Val Pro Leu Ala Asp Asp
645 650 655
Ala Ser Lys Asp Val Val Arg Thr Met Ile Arg Thr His Ile Lys Asp
660 665 670
Arg Glu Leu Arg Ser Glu Leu Met Asp Tyr Leu Asn Arg Ala Glu Thr
675 680 685
Asp Glu Glu Val Gln Glu Val Ala Asn Thr Val Asn Asp Ile Ile Asp
690 695 700
Gly Asn Ile
705
<210>36
<211>457
<212>PRT
<213>Hendra virus
<400>36
Met Asp Lys Leu Asp Leu Val Asn Asp Gly Leu Asp Ile Ile Asp Phe
1 5 10 15
Ile Gln Lys Asn Gln Lys Glu Ile Gln Lys Thr Tyr Gly Arg Ser Ser
20 25 30
Ile Gln Gln Pro Ser Thr Lys Asp Arg Thr Arg Ala Trp Glu Asp Phe
35 40 45
Leu Gln Ser Thr Ser Gly Glu His Glu Gln Ala Glu Gly Gly Met Pro
50 55 60
Lys Asn Asp Gly Gly Thr Glu Gly Arg Asn Val Glu Asp Leu Ser Ser
65 70 75 80
Val Thr Ser Ser Asp Gly Thr Ile Gly Gln Arg Val Ser Asn Thr Arg
85 90 95
Ala Trp Ala Glu Asp Pro Asp Asp Ile Gln Leu Asp Pro Met Val Thr
100 105 110
Asp Val Val Tyr His Asp His Gly Gly Glu Cys Thr Gly His Gly Pro
115 120 125
Ser Ser Ser Pro Glu Arg Gly Trp Ser Tyr His Met Ser Gly Thr His
130 135 140
Asp Gly Asn Val Arg Ala Val Pro Asp Thr Lys Val Leu Pro Asn Ala
145 150 155 160
Pro Lys Thr Thr Val Pro Glu Glu Val Arg Glu Ile Asp Leu Ile Gly
165 170 175
Leu Glu Asp Lys Phe Ala Ser Ala Gly Leu Asn Pro Ala Ala Val Pro
180 185 190
Phe Val Pro Lys Asn Gln Ser Thr Pro Thr Glu Glu Pro Pro Val Ile
195 200 205
Pro Glu Tyr Tyr Tyr Gly Ser Gly Arg Arg Gly Asp Leu Ser Lys Ser
210 215 220
Pro Pro Arg Gly Asn Val Asn Leu Asp Ser Ile Lys Ile Tyr Thr Ser
225 230 235 240
Asp Asp Glu Asp Glu Asn Gln Leu Glu Tyr Glu Asp Glu Phe Ala Lys
245 250 255
Ser Ser Ser Glu Val Val Ile Asp Thr Thr Pro Glu Asp Asn Asp Ser
260 265 270
Ile Asn Gln Glu Glu Val Val Gly Asp Pro Ser Asp Gln Gly Leu Glu
275 280 285
His Pro Phe Pro Leu Gly Lys Phe Pro Glu Lys Glu Glu Thr Pro Asp
290 295 300
Val Arg Arg Lys Asp Ser Leu Met Gln Asp Ser Cys Lys Arg Gly Gly
305 310 315 320
Val Pro Lys Arg Leu Pro Met Leu Ser Glu Glu Phe Glu Cys Ser Gly
325 330 335
Ser Asp Asp Pro Ile Ile Gln Glu Leu Glu Arg Glu Gly Ser His Pro
340 345 350
Gly Gly Ser Leu Arg Leu Arg Glu Pro Pro Gln Ser Ser Gly Asn Ser
355 360 365
Arg Asn Gln Pro Asp Arg Gln Leu Lys Thr Gly Asp Ala Ala Ser Pro
370 375 380
Gly Gly Val Gln Arg Pro Gly Thr Pro Met Pro Lys Ser Arg Ile Met
385 390 395 400
Pro Ile Lys Lys Gly His Arg Arg Glu Val Ser Ile Cys Trp Asp Gly
405 410 415
Arg Arg Ala Trp Val Glu Glu Trp Cys Asn Pro Val Cys Ser Arg Ile
420 425 430
Thr Pro Gln Pro Arg Lys Gln Glu Cys Tyr Cys Gly Glu Cys Pro Thr
435 440 445
Glu Cys Ser Gln Cys Cys His Glu Glu
450 455
<210>37
<211>166
<212>PRT
<213>Hendra virus
<400>37
Met Met Ala Ser Ile Leu Leu Thr Leu Phe Arg Arg Thr Lys Lys Lys
1 5 10 15
Tyr Lys Arg His Thr Asp Asp Gln Ala Ser Asn Asn Gln Val Pro Lys
20 25 30
Thr Gly Gln Glu His Gly Arg Thr Ser Cys Arg Ala Pro Val Glu Asn
35 40 45
Met Asn Arg Leu Arg Gly Glu Cys Leu Arg Met Met Glu Val Leu Lys
50 55 60
Glu Glu Met Trp Arg Ile Tyr Pro Val Leu Leu Pro Gln Met Glu Leu
65 70 75 80
Leu Asp Lys Glu Cys Gln Thr Pro Glu Leu Gly Gln Lys Thr Gln Met
85 90 95
Thr Tyr Asn Trp Thr Gln Trp Leu Gln Thr Leu Tyr Thr Met Ile Met
100 105 110
Glu Glu Asn Val Pro Asp Met Asp Leu Leu Gln Ala Leu Arg Glu Gly
115 120 125
Gly Val Ile Thr Cys Gln Glu His Thr Met Gly Met Tyr Val Leu Tyr
130 135 140
Leu Ile Gln Arg Cys Cys Pro Met Leu Pro Lys Leu Gln Phe Leu Lys
145 150 155 160
Lys Leu Gly Lys Leu Ile
165
<210>38
<211>352
<212>PRT
<213>Hendra virus
<400>38
Met Asp Phe Ser Val Ser Asp Asn Leu Asp Asp Pro Ile Glu Gly Val
1 5 10 15
Ser Asp Phe Ser Pro Thr Ser Trp Glu Asn Gly Gly Tyr Leu Asp Lys
20 25 30
Val Glu Pro Glu Ile Asp Lys His Gly Ser Met Ile Pro Lys Tyr Lys
35 40 45
Ile Tyr Thr Pro Gly Ala Asn Glu Arg Lys Phe Asn Asn Tyr Met Tyr
50 55 60
Met Ile Cys Tyr Gly Phe Val Glu Asp Val Glu Arg Ser Pro Glu Ser
65 70 75 80
Gly Lys Arg Lys Lys Ile Arg Thr Ile Ala Ala Tyr Pro Leu Gly Val
85 90 95
Gly Lys Ser Thr Ser His Pro Gln Asp Leu Leu Glu Glu Leu Cys Ser
100 105 110
Leu Lys Val Thr Val Arg Arg Thr Ala Gly Ala Thr Glu Lys Ile Val
115 120 125
Phe Gly Ser Ser Gly Pro Leu His His Leu Leu Pro Trp Lys Lys Ile
130 135 140
Leu Thr Gly Gly Ser Ile Phe Asn Ala Val Lys Val Cys Arg Asn Val
145 150 155 160
Asp Gln Ile Gln Leu Glu Asn Gln Gln Ser Leu Arg Ile Phe Phe Leu
165 170 175
Ser Ile Thr Lys Leu Asn Asp Ser Gly Ile Tyr Met Ile Pro Arg Thr
180 185 190
Met Leu Glu Phe Arg Arg Asn Asn Ala Ile Ala Phe Asn Leu Leu Val
195 200 205
Tyr Leu Lys Ile Asp Ala Asp Leu Ala Lys Ala Gly Ile Gln Gly Ser
210 215 220
Phe Asp Lys Asp Gly Thr Lys Val Ala Ser Phe Met Leu His Leu Gly
225 230 235 240
Asn Phe Val Arg Arg Ala Gly Lys Tyr Tyr Ser Val Glu Tyr Cys Lys
245 250 255
Arg Lys Ile Asp Arg Met Lys Leu Gln Phe Ser Leu Gly Ser Ile Gly
260 265 270
Gly Leu Ser Leu His Ile Lys Ile Asn Gly Val Ile Ser Lys Arg Leu
275 280 285
Phe Ala Gln Met Gly Phe Gln Lys Asn Leu Cys Phe Ser Leu Met Asp
290 295 300
Ile Asn Pro Trp Leu Asn Arg Leu Thr Trp Asn Asn Ser Cys Glu Ile
305 310 315 320
Ser Arg Val Ala Ala Val Leu Gln Pro Ser Val Pro Arg Glu Phe Met
325 330 335
Ile Tyr Asp Asp Val Phe Ile Asp Asn Thr Gly Lys Ile Leu Lys Gly
340 345 350
<210>39
<211>546
<212>PRT
<213>Hendra virus
<400>39
Met Ala Thr Gln Glu Val Arg Leu Lys Cys Leu Leu Cys Gly Ile Ile
1 5 10 15
Val Leu Val Leu Ser Leu Glu Gly Leu Gly Ile Leu His Tyr Glu Lys
20 25 30
Leu Ser Lys Ile Gly Leu Val Lys Gly Ile Thr Arg Lys Tyr Lys Ile
35 40 45
Lys Ser Asn Pro Leu Thr Lys Asp Ile Val Ile Lys Met Ile Pro Asn
50 55 60
Val Ser Asn Val Ser Lys Cys Thr Gly Thr Val Met Glu Asn Tyr Lys
65 70 75 80
Ser Arg Leu Thr Gly Ile Leu Ser Pro Ile Lys Gly Ala Ile Glu Leu
85 90 95
Tyr Asn Asn Asn Thr His Asp Leu Val Gly Asp Val Lys Leu Ala Gly
100 105 110
Val Val Met Ala Gly Ile Ala Ile Gly Ile Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Val Ala Leu Tyr Glu Ala Met Lys Asn Ala Asp Asn Ile
130 135 140
Asn Lys Leu Lys Ser Ser Ile Glu Ser Thr Asn Glu Ala Val Val Lys
145 150 155 160
Leu Gln Glu Thr Ala Glu Lys Thr Val Tyr Val Leu Thr Ala Leu Gln
165 170 175
Asp Tyr Ile Asn Thr Asn Leu Val Pro Thr Ile Asp Gln Ile Ser Cys
180 185 190
Lys Gln Thr Glu Leu Ala Leu Asp Leu Ala Leu Ser Lys Tyr Leu Ser
195 200 205
Asp Leu Leu Phe Val Phe Gly Pro Asn Leu Gln Asp Pro Val Ser Asn
210 215 220
Ser Met Thr Ile Gln Ala Ile Ser Gln Ala Phe Gly Gly Asn Tyr Glu
225 230 235 240
Thr Leu Leu Arg Thr Leu Gly Tyr Ala Thr Glu Asp Phe Asp Asp Leu
245 250 255
Leu Glu Ser Asp Ser Ile Ala Gly Gln Ile Val Tyr Val Asp Leu Ser
260 265 270
Ser Tyr Tyr Ile Ile Val Arg Val Tyr Phe Pro Ile Leu Thr Glu Ile
275 280 285
Gln Gln Ala Tyr Val Gln Glu Leu Leu Pro Val Ser Phe Asn Asn Asp
290 295 300
Asn Ser Glu Trp Ile Ser Ile Val Pro Asn Phe Val Leu Ile Arg Asn
305 310 315 320
Thr Leu Ile Ser Asn Ile Glu Val Lys Tyr Cys Leu Ile Thr Lys Lys
325 330 335
Ser Val Ile Cys Asn Gln Asp Tyr Ala Thr Pro Met Thr Ala Ser Val
340 345 350
Arg Glu Cys Leu Thr Gly Ser Thr Asp Lys Cys Pro Arg Glu Leu Val
355 360 365
Val Ser Ser His Val Pro Arg Phe Ala Leu Ser Gly Gly Val Leu Phe
370 375 380
Ala Asn Cys Ile Ser Val Thr Cys Gln Cys Gln Thr Thr Gly Arg Ala
385 390 395 400
Ile Ser Gln Ser Gly Glu Gln Thr Leu Leu Met Ile Asp Asn Thr Thr
405 410 415
Cys Thr Thr Val Val Leu Gly Asn Ile Ile Ile Ser Leu Gly Lys Tyr
420 425 430
Leu Gly Ser Ile Asn Tyr Asn Ser Glu Ser Ile Ala Val Gly Pro Pro
435 440 445
Val Tyr Thr Asp Lys Val Asp Ile Ser Ser Gln Ile Ser Ser Met Asn
450 455 460
Gln Ser Leu Gln Gln Ser Lys Asp Tyr Ile Lys Glu Ala Gln Lys Ile
465 470 475 480
Leu Asp Thr Val Asn Pro Ser Leu Ile Ser Met Leu Ser Met Ile Ile
485 490 495
Leu Tyr Val Leu Ser Ile Ala Ala Leu Cys Ile Gly Leu Ile Thr Phe
500 505 510
Ile Ser Phe Val Ile Val Glu Lys Lys Arg Gly Asn Tyr Ser Arg Leu
515 520 525
Asp Asp Arg Gln Val Arg Pro Val Ser Asn Gly Asp Leu Tyr Tyr Ile
530 535 540
Gly Thr
545
<210>40
<211>604
<212>PRT
<213>Hendra virus
<400>40
Met Met Ala Asp Ser Lys Leu Val Ser Leu Asn Asn Asn Leu Ser Gly
1 5 10 15
Lys Ile Lys Asp Gln Gly Lys Val Ile Lys Asn Tyr Tyr Gly Thr Met
20 25 30
Asp Ile Lys Lys Ile Asn Asp Gly Leu Leu Asp Ser Lys Ile Leu Gly
35 40 45
Ala Phe Asn Thr Val Ile Ala Leu Leu Gly Ser Ile Ile Ile Ile Val
50 55 60
Met Asn Ile Met Ile Ile Gln Asn Tyr Thr Arg Thr Thr Asp Asn Gln
65 70 75 80
Ala Leu Ile Lys Glu Ser Leu Gln Ser Val Gln Gln Gln Ile Lys Ala
85 90 95
Leu Thr Asp Lys Ile Gly Thr Glu Ile Gly Pro Lys Val Ser Leu Ile
100 105 110
Asp Thr Ser Ser Thr Ile Thr Ile Pro Ala Asn Ile Gly Leu Leu Gly
115 120 125
Ser Lys Ile Ser Gln Ser Thr Ser Ser Ile Asn Glu Asn Val Asn Asp
130 135 140
Lys Cys Lys Phe Thr Leu Pro Pro Leu Lys Ile His Glu Cys Asn Ile
145 150 155 160
Ser Cys Pro Asn Pro Leu Pro Phe Arg Glu Tyr Arg Pro Ile Ser Gln
165 170 175
Gly Val Ser Asp Leu Val Gly Leu Pro Asn Gln Ile Cys Leu Gln Lys
180 185 190
Thr Thr Ser Thr Ile Leu Lys Pro Arg Leu Ile Ser Tyr Thr Leu Pro
195 200 205
Ile Asn Thr Arg Glu Gly Val Cys Ile Thr Asp Pro Leu Leu Ala Val
210 215 220
Asp Asn Gly Phe Phe Ala Tyr Ser His Leu Glu Lys Ile Gly Ser Cys
225 230 235 240
Thr Arg Gly Ile Ala Lys Gln Arg Ile Ile Gly Val Gly Glu Val Leu
245 250 255
Asp Arg Gly Asp Lys Val Pro Ser Met Phe Met Thr Asn Val Trp Thr
260 265 270
Pro Pro Asn Pro Ser Thr Ile His His Cys Ser Ser Thr Tyr His Glu
275 280 285
Asp Phe Tyr Tyr Thr Leu Cys Ala Val Ser His Val Gly Asp Pro Ile
290 295 300
Leu Asn Ser Thr Ser Trp Thr Glu Ser Leu Ser Leu Ile Arg Leu Ala
305 310 315 320
Val Arg Pro Lys Ser Asp Ser Gly Asp Tyr Asn Gln Lys Tyr Ile Ala
325 330 335
Ile Thr Lys Val Glu Arg Gly Lys Tyr Asp Lys Val Met Pro Tyr Gly
340 345 350
Pro Ser Gly Ile Lys Gln Gly Asp Thr Leu Tyr Phe Pro Ala Val Gly
355 360 365
Phe Leu Pro Arg Thr Glu Phe Gln Tyr Asn Asp Ser Asn Cys Pro Ile
370 375 380
Ile His Cys Lys Tyr Ser Lys Ala Glu Asn Cys Arg Leu Ser Met Gly
385 390 395 400
Val Asn Ser Lys Ser His Tyr Ile Leu Arg Ser Gly Leu Leu Lys Tyr
405 410 415
Asn Leu Ser Leu Gly Gly Asp Ile Ile Leu Gln Phe Ile Glu Ile Ala
420 425 430
Asp Asn Arg Leu Thr Ile Gly Ser Pro Ser Lys Ile Tyr Asn Ser Leu
435 440 445
Gly Gln Pro Val Phe Tyr Gln Ala Ser Tyr Ser Trp Asp Thr Met Ile
450 455 460
Lys Leu Gly Asp Val Asp Thr Val Asp Pro Leu Arg Val Gln Trp Arg
465 470 475 480
Asn Asn Ser Val Ile Ser Arg Pro Gly Gln Ser Gln Cys Pro Arg Phe
485 490 495
Asn Val Cys Pro Glu Val Cys Trp Glu Gly Thr Tyr Asn Asp Ala Phe
500 505 510
Leu Ile Asp Arg Leu Asn Trp Val Ser Ala Gly Val Tyr Leu Asn Ser
515 520 525
Asn Gln Thr Ala Glu Asn Pro Val Phe Ala Val Phe Lys Asp Asn Glu
530 535 540
Ile Leu Tyr Gln Val Pro Leu Ala Glu Asp Asp Thr Asn Ala Gln Lys
545 550 555 560
Thr Ile Thr Asp Cys Phe Leu Leu Glu Asn Val Ile Trp Cys Ile Ser
565 570 575
Leu Val Glu Ile Tyr Asp Thr Gly Asp Ser Val Ile Arg Pro Lys Leu
580 585 590
Phe Ala Val Lys Ile Pro Ala Gln Cys Ser Glu Ser
595 600
<210>41
<211>2244
<212>PRT
<213>Hendra virus
<400>41
Met Ala His Glu Leu Ser Ile Ser Asp Ile Ile Tyr Pro Glu Cys His
1 5 10 15
Leu Asp Ser Pro Ile Val Ser Gly Lys Leu Ile Ser Ala Ile Glu Tyr
20 25 30
Ala Gln Leu Arg His Asn Gln Pro Asn Gly Asp Lys Arg Leu Thr Glu
35 40 45
Asn Ile Lys Ile Asn Leu Gln Gly Lys Arg Arg Ser Val Tyr Ile Ser
50 55 60
Arg Gln Ser Arg Leu Gly Asn Tyr Ile Arg Asp Asn Ile Lys Asn Leu
65 70 75 80
Lys Glu Phe Leu His Val Ser Tyr Pro Glu Cys Asn Lys Ser Leu Phe
85 90 95
Ser Leu Lys Ser Pro Gly Met Thr Ser Lys Leu Ser Asn Ile Met Lys
100 105 110
Lys Ser Phe Lys Ala Tyr Asn Ile Val Ser Arg Lys Ile Ile Glu Met
115 120 125
Leu Gln Asn Ile Thr Arg Asn Leu Ile Thr Gln Asp Gln Lys Asp Glu
130 135 140
Val Leu Gly Ile Tyr Glu Gln Asp Arg Leu Ser Asn Ile Gly Lys Tyr
145 150 155 160
Met Ser Gln Ser Gln Trp Tyr Glu Cys Phe Leu Phe Trp Phe Thr Ile
165 170 175
Lys Thr Glu Met Arg Ala Val Ile Lys Asn Ser Gln Lys Pro Lys Phe
180 185 190
Arg Ser Asp Ser Cys Ile Ile His Met Lys Asp Asn Asn Met Glu Ile
195 200 205
Val Met Asn Pro Asn Leu Val Cys Ile Tyr Lys Asn Asp Lys Asp Gly
210 215 220
Lys Arg Cys Tyr Tyr Leu Thr Pro Glu Ile Val Leu Met Cys Cys Asp
225 230 235 240
Val Leu Glu Gly Arg Met Met Ile Glu Thr Ser Ile Lys Ser Asp Ile
245 250 255
Lys Tyr Gln Ser Leu Ile Thr Arg Ser Asn Ala Leu Trp Thr Phe Ile
260 265 270
Asp Ser Leu Phe Pro Ile Met Gly Asn Arg Ile Tyr Asn Ile Val Ser
275 280 285
Met Ile Glu Pro Leu Val Leu Ala Leu Leu Gln Leu Lys Asp Glu Ala
290 295 300
Arg Ile Leu Arg Gly Ala Phe Leu His His Cys Ile Lys Glu Ile His
305 310 315 320
Gln Glu Leu Ile Gly Cys Gly Phe Thr Asp Gln Lys Thr Arg Ser Ile
325 330 335
Phe Ile Asp Asp Leu Leu Ser Val Met Asn Ile Asp Asn Ile His Leu
340 345 350
Leu Ala Glu Phe Phe Ser Phe Phe Arg Thr Phe Gly His Pro Ile Leu
355 360 365
Glu Ala Lys Thr Ala Ala Asp Lys Val Arg Glu His Met Leu Ala Asp
370 375 380
Lys Val Leu Glu Tyr Gly Pro Ile Met Lys Ala His Ala Val Phe Cys
385 390 395 400
Gly Thr Ile Ile Asn Gly Tyr Arg Asp Arg His Arg Gly Ala Trp Pro
405 410 415
Pro Leu Tyr Leu Pro Ser His Ala Ser Lys His Ile Ile Arg Leu Lys
420 425 430
Asn Ser Gly Glu Ser Leu Thr Val Asp Asp Cys Val Lys Asn Trp Glu
435 440 445
Ser Phe Cys Gly Ile Gln Phe Asp Cys Phe Met Glu Leu Lys Leu Asp
450 455 460
Ser Asp Leu Ser Met Tyr Met Lys Asp Lys Ala Leu Ser Pro Ile Lys
465 470 475 480
Glu Glu Trp Asp Ser Val Tyr Pro Arg Glu Val Leu Asn Tyr Thr Pro
485 490 495
Pro Arg Ser Thr Glu Pro Arg Arg Leu Val Asp Val Phe Val Asn Asp
500 505 510
Glu Asn Phe Asp Pro Tyr Asn Met Leu Glu Tyr Val Leu Thr Gly Asp
515 520 525
Tyr Leu Thr Asp Glu Gln Phe Asn Val Ser Tyr Ser Leu Lys Glu Lys
530 535 540
Glu Thr Lys Gln Ala Gly Arg Leu Phe Ala Lys Met Thr Tyr Lys Met
545 550 555 560
Arg Ala Cys Gln Val Ile Ala Glu Ala Leu Ile Ala Ser Gly Val Gly
565 570 575
Lys Tyr Phe Lys Glu Asn Gly Met Val Lys Asp Glu His Glu Leu Leu
580 585 590
Lys Thr Leu Phe Gln Leu Ser Ile Ser Ser Val Pro Arg Gly Asn Ser
595 600 605
Gln Gly Arg Asp Ser Glu Phe Ser Asn Asn Thr Glu Lys Ser Leu Ile
610 615 620
Ser Leu Lys Arg Thr Thr Gly Arg Leu Leu Asn Asn Glu Val Pro Cys
625 630 635 640
Arg Met Asn Ile Met Ser Ala Leu Ile Asp Lys Asn Gln Ser Asp Gln
645 650 655
Lys Lys His Asn Ile Leu Pro Asn Thr Arg Asn Arg His Lys Cys Asp
660 665 670
Asn Thr Ser Gln Thr Phe Leu Asp Tyr His Met Glu Phe Ser Pro Tyr
675 680 685
Lys Ser Asp Arg Met Asp Arg Thr Glu Thr Ser Asp Phe Ser Lys Tyr
690 695 700
Asp Asp Gly Thr Gly Thr Lys Phe Asp Thr Val Ser Ala Phe Leu Thr
705 710 715 720
Thr Asp Leu Lys Lys Phe Cys Leu Asn Trp Arg Tyr Glu Ser Met Ala
725 730 735
Ile Phe Ala Glu Arg Leu Asp Glu Ile Tyr Gly Leu Pro Gly Phe Phe
740 745 750
Asn Trp Met His Lys Arg Leu Glu Lys Ser Val Ile Tyr Val Ala Asp
755 760 765
Pro Asn Cys Pro Pro Asp Ile Gly Lys His Ile Asn Leu Asp Asp Thr
770 775 780
Pro Glu Asp Asp Ile Phe Ile His Ser Pro Lys Gly Gly Ile Glu Gly
785 790 795 800
Tyr Ser Gln Lys Thr Trp Thr Ile Ala Thr Ile Pro Phe Leu Phe Leu
805 810 815
Ser Ala Tyr Glu Thr Asn Thr Arg Ile Ala Ala Ile Val Gln Gly Asp
820 825 830
Asn Glu Ser Ile Ala Ile Thr Gln Lys Val His Pro Asn Leu Pro Tyr
835 840 845
Lys Val Lys Lys Glu Ile Cys Ala Arg Gln Ala Gln Leu Tyr Phe Asp
850 855 860
Arg Leu Arg Met Asn Leu Arg Ala Leu Gly Leu Asn Leu Lys Ala Thr
865 870 875 880
Glu Thr Ile Ile Ser Thr His Leu Phe Val Tyr Ser Lys Lys Ile His
885 890 895
Tyr Asp Gly Ala Val Leu Ser Gln Ala Leu Lys Ser Met Ser Arg Cys
900 905 910
Cys Phe Trp Ser Glu Thr Leu Val Asp Glu Thr Arg Ser Ala Cys Ser
915 920 925
Asn Ile Ser Thr Thr Ile Ala Lys Ala Ile Glu Asn Gly Leu Ser Arg
930 935 940
Asn Val Gly Tyr Cys Ile Asn Val Leu Lys Val Ile Gln Gln Leu Leu
945 950 955 960
Ile Ser Thr Glu Phe Ser Ile Asn Glu Thr Leu Thr Ala Asp Val Thr
965 970 975
Ser Pro Ile Ser Asn Asn Leu Asp Trp Leu Val Thr Ala Ser Arg Ile
980 985 990
Pro Ala Pro Ile Gly Gly Phe Asn Tyr Leu Asn Leu Ser Arg Ile Phe
995 1000 1005
Val Arg Asn Ile Gly Asp Pro Val Thr Ala Ser Leu Ala Asp Leu
1010 1015 1020
Lys Arg Met Ile Glu His Asp Leu Met Thr Asp Lys Val Leu Gln
1025 1030 1035
Lys Val Met Asn Gln Glu Pro Gly Asp Ala Ser Phe Leu Asp Trp
1040 1045 1050
Ala Ser Asp Pro Tyr Ser Gly Asn Leu Pro Asp Ser Gln Ser Ile
1055 1060 1065
Thr Lys Thr Ile Lys Asn Ile Thr Ala Arg Thr Ile Leu Arg Thr
1070 1075 1080
Ser Pro Asn Pro Met Leu Lys Gly Leu Phe His Asp Lys Ser Phe
1085 1090 1095
Glu Glu Asp Leu Glu Leu Ala Thr Phe Leu Met Asp Arg Arg Ile
1100 1105 1110
Ile Leu Pro Arg Ala Ala His Glu Ile Leu Asp Asn Ser Leu Thr
1115 1120 1125
Gly Ala Arg Glu Glu Ile Ala Gly Leu Leu Asp Thr Thr Lys Gly
1130 1135 1140
Leu Ile Arg Ser Gly Leu Lys Lys Ser Gly Ile Gln Pro Lys Leu
1145 1150 1155
Val Ser Arg Leu Ser Asn His Asp Tyr Asn Gln Phe Leu Ile Leu
1160 1165 1170
Asn Arg Leu Leu Ser Asn Lys Lys Arg Asn Asp Leu Ile Ser Pro
1175 1180 1185
Lys Thr Cys Ser Val Asp Leu Ala Lys Ala Leu Arg Cys His Met
1190 1195 1200
Trp Arg Asp Leu Ala Leu Gly Arg Ser Ile Tyr Gly Leu Glu Val
1205 1210 1215
Pro Asp Ala Leu Glu Ala Met Thr Gly Arg Tyr Ile Thr Gly Ser
1220 1225 1230
Met Glu Cys Gln Leu Cys Asp Gln Gly Asn Thr Met Tyr Gly Trp
1235 1240 1245
Phe Phe Val Pro Arg Asp Ser Gln Leu Asp Gln Val Asn Lys Glu
1250 1255 1260
His Ser Ser Ile Arg Val Pro Tyr Val Gly Ser Ser Thr Asp Glu
1265 1270 1275
Arg Ser Asp Ile Lys Leu Gly Asn Val Lys Arg Pro Thr Arg Ala
1280 1285 1290
Leu Arg Ser Ala Ile Glu Leu Gln Leu Phe Ile Pro Gly His Thr
1295 1300 1305
Glu Ile Leu Lys Lys Val Gly Met Arg Leu Gly Thr Trp Leu Pro
1310 1315 1320
Arg Gly Val Asn Ile Asp Ile Asp Val Leu Lys Ala Ile Thr Pro
1325 1330 1335
Val Ser Thr Ser Asn Asn Leu Ser His Arg Leu Arg Asp Arg Ser
1340 1345 1350
Thr Gln Phe Lys Leu Pro Gly Ser Val Leu Asn Arg Val Ser Arg
1355 1360 1365
Tyr Val Asn Ile Ser Asn Asp Asn Leu Asp Phe Arg Val Glu Gly
1370 1375 1380
Glu Lys Val Asp Thr Asn Leu Ile Tyr Gln Gln Thr Met Leu Leu
1385 1390 1395
Gly Leu Ser Val Leu Glu Gly Lys Phe Arg Leu Arg Thr Glu Thr
1400 1405 1410
Asp Asp Tyr Asn Gly Ile Tyr His Leu His Val Arg Asp Asn Cys
1415 1420 1425
Cys Val Lys Glu Val Ala Asp Ile Gly Gly Val Asn Ala Glu Leu
1430 1435 1440
Pro Val Pro Glu Tyr Thr Glu Val Glu Asn Asn Arg Leu Ile Tyr
1445 1450 1455
Asp Pro Asp Pro Val Ser Glu Ile Asp Cys Asp Arg Leu Ser Lys
1460 1465 1470
Gln Glu Ser Lys Ala Arg Glu Leu Asp Phe Pro Leu Trp Ser Thr
1475 1480 1485
Glu Glu Leu His Asp Val Leu Ala Lys Thr Val Ala Gln Thr Val
1490 1495 1500
Leu Glu Ile Ile Thr Lys Ala Asp Lys Asp Val Leu Lys Gln His
1505 1510 1515
Leu Ala Ile Asp Ser Asp Asp Ser Ile Asn Ser Leu Ile Thr Glu
1520 1525 1530
Phe Leu Met Val Asp Pro Glu Leu Phe Ala Leu Tyr Leu Gly Gln
1535 1540 1545
Ser Ile Ser Val Lys Trp Ala Phe Glu Ile His His Arg Arg Pro
1550 1555 1560
His Gly Arg His Thr Met Val Asp Leu Leu Ser Asp Leu Ile Ser
1565 1570 1575
Asn Thr Ser Lys His Thr Tyr Lys Val Leu Ser Asn Ala Leu Ser
1580 1585 1590
His Pro Arg Val Phe Lys Arg Phe Val Asn Cys Gly Leu Leu Leu
1595 1600 1605
Pro Thr Gln Gly Pro Tyr Leu His Gln Gln Asp Phe Glu Lys Leu
1610 1615 1620
Ser Gln Asn Leu Leu Ile Thr Ser Tyr Met Asn Tyr Leu Met Asn
1625 1630 1635
Trp Cys Asp Phe Lys Lys Phe Pro Phe Leu Ile Ala Glu Gln Asp
1640 1645 1650
Glu Ala Val Val Glu Leu Arg Glu Asp Ile Ile Thr Ser Lys His
1655 1660 1665
Leu Cys Met Ile Ile Asp Leu Tyr Ala Asn His His Lys Pro Pro
1670 1675 1680
Trp Ile Ile Asp Leu Asn Pro Gln Glu Lys Ile Cys Val Leu Arg
1685 1690 1695
Asp Phe Ile Ser Lys Cys Arg His Thr Asp Val Ser Ser Arg Ser
1700 1705 1710
Trp Asn Ile Thr Asp Leu Asp Phe Met Val Phe Tyr Ala Ser Leu
1715 1720 1725
Thr Tyr Leu Arg Arg Gly Ile Ile Lys Gln Leu Arg Ile Arg Gln
1730 1735 1740
Val Thr Glu Val Ile Asp Thr Thr Thr Met Leu Arg Asp Asn Ile
1745 1750 1755
Leu Val Glu Asn Pro Pro Ile Lys Thr Gly Val Leu Asp Ile Arg
1760 1765 1770
Gly Cys Ile Ile Tyr Asn Leu Glu Glu Ile Leu Ser Met Asn Thr
1775 1780 1785
Lys Ser Thr Ser Arg Lys Val Phe Asn Leu Gly Ser Lys Leu Ser
1790 1795 1800
Val Glu Asn His Lys Tyr Arg Arg Ile Gly Leu Asn Ser Ser Ser
1805 1810 1815
Cys Tyr Lys Ala Leu Asn Leu Ser Pro Leu Ile Gln Arg Tyr Leu
1820 1825 1830
Pro Ala Gly Ser Gln Arg Leu Phe Val Gly Glu Gly Ser Gly Ser
1835 1840 1845
Met Met Leu Leu Tyr Gln Gln Thr Leu Gly Cys Ser Ile Ser Phe
1850 1855 1860
Tyr Asn Ser Gly Ile Asp Gly Asp Tyr Ile Pro Gly Gln Arg Glu
1865 1870 1875
Leu Arg Leu Phe Pro Ser Glu Tyr Ser Ile Ala Glu Asp Asp Pro
1880 1885 1890
Ser Gln Ser Asp Lys Leu Lys Gly Leu Val Val Pro Leu Phe Asn
1895 1900 1905
Gly Arg Pro Glu Thr Thr Trp Ile Gly Asn Leu Asp Ser Tyr Glu
1910 1915 1920
Tyr Ile Ile Asn Arg Thr Ala Gly Arg Asn Ile Gly Leu Val His
1925 1930 1935
Ser Asp Met Glu Ser Gly Ile Asp Lys Gln Val Glu Glu Ile Met
1940 1945 1950
Ile Glu His Ser His Leu Ile Ser Ile Ala Ile Asn Val Met Ile
1955 1960 1965
Glu Asp Gly Val Leu Val Ser Lys Ile Ala Phe Ala Pro Gly Phe
1970 1975 1980
Pro Ile Ser Arg Leu Leu Asn Met Tyr Arg Ser Tyr Phe Gly Leu
1985 1990 1995
Val Leu Val Cys Phe Pro Val Tyr Ser Asn Pro Glu Ser Thr Glu
2000 2005 2010
Val Tyr Leu Ile Cys Leu Gln Lys Thr Ile Lys Thr Ile Ile Pro
2015 2020 2025
Pro Gln Lys Val Leu Asp His Ser Tyr Leu Ser Asp Glu Ile Asn
2030 2035 2040
Asp Gln Gly Ile Thr Ser Val Ile Phe Lys Ile Lys Asn Ile Gln
2045 2050 2055
Ser Lys Gln Phe His Glu Asp Leu Val Lys His Tyr Gln Val Glu
2060 2065 2070
Gln Pro Phe Phe Val Pro Ser His Ile Thr Cys Asp Glu Lys Leu
2075 2080 2085
Leu Met Gln Ala Gly Leu Lys Met Asn Gly Pro Glu Ile Leu Lys
2090 2095 2100
Asn Glu Val Gly Tyr Asp Ile Gly Ser Asp Ile Asn Thr Leu Arg
2105 2110 2115
Ser Thr Ile Ile Ile Leu Leu Asn Glu Ala Met Asn Tyr Phe Asp
2120 2125 2130
Asp Glu Arg Ser Pro Ser His His Leu Glu Pro Phe Pro Val Leu
2135 2140 2145
Glu Lys Thr Arg Val Lys Thr Ile Met Gly Arg Val Thr Arg Lys
2150 2155 2160
Val Thr Val Tyr Ser Leu Ile Lys Leu Lys Glu Thr Lys Ser Pro
2165 2170 2175
Glu Leu Tyr Asn Ile Lys Asn Tyr Ile Arg Arg Lys Val Leu Ile
2180 2185 2190
Leu Asp Phe Arg Ser His Thr Met Ile Lys Leu Leu Pro Lys Gly
2195 2200 2205
Met Lys Glu Arg Arg Glu Lys Ser Gly Phe Lys Glu Ile Trp Ile
2210 2215 2220
Phe Asp Leu Ser Asn Arg Glu Val Lys Ile Trp Trp Lys Ile Ile
2225 2230 2235
Gly Tyr Leu Ser Leu Val
2240
<210>42
<211>707
<212>PRT
<213>Hendra virus
<400>42
Met Asp Lys Leu Asp Leu Val Asn Asp Gly Leu Asp Ile Ile Asp Phe
1 5 10 15
Ile Gln Lys Asn Gln Lys Glu Ile Gln Lys Thr Tyr Gly Arg Ser Ser
20 25 30
Ile Gln Gln Pro Ser Thr Lys Asp Arg Thr Arg Ala Trp Glu Asp Phe
35 40 45
Leu Gln Ser Thr Ser Gly Glu His Glu Gln Ala Glu Gly Gly Met Pro
50 55 60
Lys Asn Asp Gly Gly Thr Glu Gly Arg Asn Val Glu Asp Leu Ser Ser
65 70 75 80
Val Thr Ser Ser Asp Gly Thr Ile Gly Gln Arg Val Ser Asn Thr Arg
85 90 95
Ala Trp Ala Glu Asp Pro Asp Asp Ile Gln Leu Asp Pro Met Val Thr
100 105 110
Asp Val Val Tyr His Asp His Gly Gly Glu Cys Thr Gly His Gly Pro
115 120 125
Ser Ser Ser Pro Glu Arg Gly Trp Ser Tyr His Met Ser Gly Thr His
130 135 140
Asp Gly Asn Val Arg Ala Val Pro Asp Thr Lys Val Leu Pro Asn Ala
145 150 155 160
Pro Lys Thr Thr Val Pro Glu Glu Val Arg Glu Ile Asp Leu Ile Gly
165 170 175
Leu Glu Asp Lys Phe Ala Ser Ala Gly Leu Asn Pro Ala Ala Val Pro
180 185 190
Phe Val Pro Lys Asn Gln Ser Thr Pro Thr Glu Glu Pro Pro Val Ile
195 200 205
Pro Glu Tyr Tyr Tyr Gly Ser Gly Arg Arg Gly Asp Leu Ser Lys Ser
210 215 220
Pro Pro Arg Gly Asn Val Asn Leu Asp Ser Ile Lys Ile Tyr Thr Ser
225 230 235 240
Asp Asp Glu Asp Glu Asn Gln Leu Glu Tyr Glu Asp Glu Phe Ala Lys
245 250 255
Ser Ser Ser Glu Val Val Ile Asp Thr Thr Pro Glu Asp Asn Asp Ser
260 265 270
Ile Asn Gln Glu Glu Val Val Gly Asp Pro Ser Asp Gln Gly Leu Glu
275 280 285
His Pro Phe Pro Leu Gly Lys Phe Pro Glu Lys Glu Glu Thr Pro Asp
290 295 300
Val Arg Arg Lys Asp Ser Leu Met Gln Asp Ser Cys Lys Arg Gly Gly
305 310 315 320
Val Pro Lys Arg Leu Pro Met Leu Ser Glu Glu Phe Glu Cys Ser Gly
325 330 335
Ser Asp Asp Pro Ile Ile Gln Glu Leu Glu Arg Glu Gly Ser His Pro
340 345 350
Gly Gly Ser Leu Arg Leu Arg Glu Pro Pro Gln Ser Ser Gly Asn Ser
355 360 365
Arg Asn Gln Pro Asp Arg Gln Leu Lys Thr Gly Asp Ala Ala Ser Pro
370 375 380
Gly Gly Val Gln Arg Pro Gly Thr Pro Met Pro Lys Ser Arg Ile Met
385 390 395 400
Pro Ile Lys Lys Gly Thr Asp Ala Lys Ser Gln Tyr Val Gly Thr Glu
405 410 415
Asp Val Pro Gly Ser Lys Ser Gly Ala Thr Arg Tyr Val Arg Gly Leu
420 425 430
Pro Pro Asn Gln Glu Ser Lys Ser Val Thr Ala Glu Asn Val Gln Leu
435 440 445
Ser Ala Pro Ser Ala Val Thr Arg Asn Glu Gly His Asp Gln Glu Val
450 455 460
Thr Ser Asn Glu Asp Ser Leu Asp Asp Asn Ile Leu Cys His Gln Met
465 470 475 480
Ile Leu Leu Thr Leu Phe Leu Pro His Asp Thr Asp Arg Leu Asn Tyr
485 490 495
His Ala Asp His Leu Asn Asp Tyr Asp Leu Glu Thr Leu Cys Glu Glu
500 505 510
Ser Val Leu Met Gly Ile Val Asn Ala Ile Lys Leu Ile Asn Ile Asp
515 520 525
Met Arg Leu Asn His Ile Glu Glu Gln Met Lys Glu Ile Pro Lys Ile
530 535 540
Ile Asn Lys Ile Asp Ser Ile Asp Arg Val Leu Ala Lys Thr Asn Thr
545 550 555 560
Ala Leu Ser Thr Ile Glu Gly His Leu Val Ser Met Met Ile Met Ile
565 570 575
Pro Gly Lys Gly Lys Gly Glu Arg Lys Gly Lys Thr Asn Pro Glu Leu
580 585 590
Lys Pro Val Ile Gly Arg Asn Ile Leu Glu Gln Gln Glu Leu Phe Ser
595 600 605
Phe Asp Asn Leu Lys Asn Phe Arg Asp Gly Ser Leu Thr Asp Glu Pro
610 615 620
Tyr Gly Gly Val Ala Arg Ile Arg Asp Asp Leu Ile Leu Pro Glu Leu
625 630 635 640
Asn Phe Ser Glu Thr Asn Ala Ser Gln Phe Val Pro Leu Ala Asp Asp
645 650 655
Ala Ser Lys Asp Val Val Arg Thr Met Ile Arg Thr His Ile Lys Asp
660 665 670
Arg Glu Leu Arg Ser Glu Leu Met Asp Tyr Leu Asn Arg Ala Glu Thr
675 680 685
Asp Glu Glu Val Gln Glu Val Ala Asn Thr Val Asn Asp Ile Ile Asp
690 695 700
Gly Asn Ile
705
<210>43
<211>457
<212>PRT
<213>Hendra virus
<400>43
Met Asp Lys Leu Asp Leu Val Asn Asp Gly Leu Asp Ile Ile Asp Phe
1 5 10 15
Ile Gln Lys Asn Gln Lys Glu Ile Gln Lys Thr Tyr Gly Arg Ser Ser
20 25 30
Ile Gln Gln Pro Ser Thr Lys Asp Arg Thr Arg Ala Trp Glu Asp Phe
35 40 45
Leu Gln Ser Thr Ser Gly Glu His Glu Gln Ala Glu Gly Gly Met Pro
50 55 60
Lys Asn Asp Gly Gly Thr Glu Gly Arg Asn Val Glu Asp Leu Ser Ser
65 70 75 80
Val Thr Ser Ser Asp Gly Thr Ile Gly Gln Arg Val Ser Asn Thr Arg
85 90 95
Ala Trp Ala Glu Asp Pro Asp Asp Ile Gln Leu Asp Pro Met Val Thr
100 105 110
Asp Val Val Tyr His Asp His Gly Gly Glu Cys Thr Gly His Gly Pro
115 120 125
Ser Ser Ser Pro Glu Arg Gly Trp Ser Tyr His Met Ser Gly Thr His
130 135 140
Asp Gly Asn Val Arg Ala Val Pro Asp Thr Lys Val Leu Pro Asn Ala
145 150 155 160
Pro Lys Thr Thr Val Pro Glu Glu Val Arg Glu Ile Asp Leu Ile Gly
165 170 175
Leu Glu Asp Lys Phe Ala Ser Ala Gly Leu Asn Pro Ala Ala Val Pro
180 185 190
Phe Val Pro Lys Asn Gln Ser Thr Pro Thr Glu Glu Pro Pro Val Ile
195 200 205
Pro Glu Tyr Tyr Tyr Gly Ser Gly Arg Arg Gly Asp Leu Ser Lys Ser
210 215 220
Pro Pro Arg Gly Asn Val Asn Leu Asp Ser Ile Lys Ile Tyr Thr Ser
225 230 235 240
Asp Asp Glu Asp Glu Asn Gln Leu Glu Tyr Glu Asp Glu Phe Ala Lys
245 250 255
Ser Ser Ser Glu Val Val Ile Asp Thr Thr Pro Glu Asp Asn Asp Ser
260 265 270
Ile Asn Gln Glu Glu Val Val Gly Asp Pro Ser Asp Gln Gly Leu Glu
275 280 285
His Pro Phe Pro Leu Gly Lys Phe Pro Glu Lys Glu Glu Thr Pro Asp
290 295 300
Val Arg Arg Lys Asp Ser Leu Met Gln Asp Ser Cys Lys Arg Gly Gly
305 310 315 320
Val Pro Lys Arg Leu Pro Met Leu Ser Glu Glu Phe Glu Cys Ser Gly
325 330 335
Ser Asp Asp Pro Ile Ile Gln Glu Leu Glu Arg Glu Gly Ser His Pro
340 345 350
Gly Gly Ser Leu Arg Leu Arg Glu Pro Pro Gln Ser Ser Gly Asn Ser
355 360 365
Arg Asn Gln Pro Asp Arg Gln Leu Lys Thr Gly Asp Ala Ala Ser Pro
370 375 380
Gly Gly Val Gln Arg Pro Gly Thr Pro Met Pro Lys Ser Arg Ile Met
385 390 395 400
Pro Ile Lys Lys Gly His Arg Arg Glu Val Ser Ile Cys Trp Asp Gly
405 410 415
Arg Arg Ala Trp Val Glu Glu Trp Cys Asn Pro Val Cys Ser Arg Ile
420 425 430
Thr Pro Gln Pro Arg Lys Gln Glu Cys Tyr Cys Gly Glu Cys Pro Thr
435 440 445
Glu Cys Ser Gln Cys Cys His Glu Glu
450 455
<210>44
<211>166
<212>PRT
<213>Hendra virus
<400>44
Met Met Ala Ser Ile Leu Leu Thr Leu Phe Arg Arg Thr Lys Lys Lys
1 5 10 15
Tyr Lys Arg His Thr Asp Asp Gln Ala Ser Asn Asn Gln Val Pro Lys
20 25 30
Thr Gly Gln Glu His Gly Arg Thr Ser Cys Arg Ala Pro Val Glu Asn
35 40 45
Met Asn Arg Leu Arg Gly Glu Cys Leu Arg Met Met Glu Val Leu Lys
50 55 60
Glu Glu Met Trp Arg Ile Tyr Pro Val Leu Leu Pro Gln Met Glu Leu
65 70 75 80
Leu Asp Lys Glu Cys Gln Thr Pro Glu Leu Gly Gln Lys Thr Gln Met
85 90 95
Thr Tyr Asn Trp Thr Gln Trp Leu Gln Thr Leu Tyr Thr Met Ile Met
100 105 110
Glu Glu Asn Val Pro Asp Met Asp Leu Leu Gln Ala Leu Arg Glu Gly
115 120 125
Gly Val Ile Thr Cys Gln Glu His Thr Met Gly Met Tyr Val Leu Tyr
130 135 140
Leu Ile Gln Arg Cys Cys Pro Met Leu Pro Lys Leu Gln Phe Leu Lys
145 150 155 160
Lys Leu Gly Lys Leu Ile
165
<210>45
<211>2698
<212>DNA
<213>Hendra virus
<400>45
aggatccaag accataaatc taggatcctt tacaatctca gaccctggtg caaggttata 60
tatagagccc aattctccta attaaacagt gtctcaggtt gacaaatgga caagttggat 120
ctagtcaatg atggcctcga tattattgac tttattcaga agaaccaaaa agaaatacaa 180
aagacatacg gacgatcaag catccaacaa ccaagtacca aagacaggac aagagcatgg 240
gaggacttct tgcagagcac cagtggagaa catgaacagg ctgagggggg aatgcctaag 300
aatgatggag gtactgaagg aagaaatgtg gaggatctat ccagtgttac ttcctcagat 360
ggaactattg gacaaagagt gtcaaacacc cgagcttggg cagaagaccc agatgacata 420
caactggacc caatggttac agacgttgta taccatgatc atggaggaga atgtaccgga 480
catggacctt cttcaagccc tgagagaggg tggagttatc acatgtcagg aacacacgat 540
gggaatgtac gtgctgtacc tgatacaaag gtgttgccca atgctcccaa aactacagtt 600
cctgaagaag ttagggaaat tgatttaatt gggttggagg acaaatttgc atcagcggga 660
ttaaatccag ctgcagttcc ctttgtcccc aagaatcaat caaccccaac tgaggagcct 720
ccagtcatcc cggagtatta ctatggatct gggaggagag gagatctctc aaaatctcct 780
cctagaggta atgtcaatct ggacagcatc aagatttaca cttcggacga cgaggatgaa 840
aatcagctgg agtatgagga tgagtttgcc aaaagctcaa gtgaggttgt cattgacact 900
actcctgagg acaatgattc tatcaaccag gaggaagtag ttggggaccc gtctgatcag 960
ggtttagagc atcctttccc tttggggaaa ttcccggaga aagaagaaac tcctgatgta 1020
cgcagaaagg attccctaat gcaggacagt tgtaagagag gaggagtacc gaaaagactg 1080
cccatgttat ctgaagagtt cgagtgctcc ggatcagatg atccaataat acaagaatta 1140
gaacgagaag ggtctcatcc aggaggttca ttacgtttga gggaaccacc tcagtcgtca 1200
gggaattcca gaaatcaacc tgaccgtcag ctgaagacag gtgatgcagc atcacccgga 1260
ggcgtccaga gaccaggcac accgatgcct aaatcaagga tcatgcccat taaaaagggc 1320
acagacgcga agtctcaata tgttgggacg gaagacgtgc ctgggtcgaa gagtggtgca 1380
acccggtatg ttcgcggatt accccccaac caagaaagca agagtgttac tgcggagaat 1440
gtccaactga gtgctcccag tgctgtcacg aggaatgaag gacacgacca ggaagttacc 1500
agcaacgaag atagtctgga tgacaatata ttatgccatc agatgatttt gctaacactt 1560
ttcttacccc atgacactga taggttaaat tatcatgcag accatctcaa tgattacgat 1620
ttagagacct tgtgtgagga gtcagtattg atgggcatcg ttaacgcaat caaactcatc 1680
aacattgata tgaggttgaa tcacattgag gaacagatga aagagatacc caagatcatc 1740
aacaaaatag actccattga tcgagtgttg gctaagacca atactgcatt gtcgacaata 1800
gaaggacacc tagtctcgat gatgatcatg ataccaggga aaggcaaggg tgaaaggaaa 1860
gggaaaacaa atccggagct aaaacctgta attgggagaa atatcctaga acagcaagag 1920
ttattttcat tcgacaatct caagaatttc agagatgggt cattgactga tgagccctat 1980
ggaggggtgg cccgaataag agatgatttg atcctgcctg aactcaactt cagtgagaca 2040
aatgcatcac aatttgttcc tttggcagat gatgcatcta aggatgtcgt gagaactatg 2100
attaggactc acatcaagga cagagagttg aggtctgagt tgatggatta tctaaatcga 2160
gcagaaacag atgaagaggt tcaggaggtg gccaatacag tcaatgatat tattgatggg 2220
aacatctaaa catgtcagta atttgataga gtagcagtcg aaccaagatc atatattaga 2280
ggtacacatc ataatggcca gtgtcttaat ctcaaccatt cataaaccgt cagtgtttcc 2340
cttataaact ccattatgta gtttattaat atctagagct attctttatt aagtaataaa 2400
tctgtattag attattatat aatctctata atcaacacgc ctattacttc atgcacttac 2460
tatcaccatt taataatagt tcagatcacc ccttcaaaat ctgggttgtg ccaagtcatt 2520
ttcctcacca tataatccac gcagtatgca ttcaaatact aaaccacatt cagacagagt 2580
tagacattca agcaatcaat aactatatta actataagaa tgatgaatca gccctgcgcc 2640
aacaaggcaa tgtcaatcac ttgaatgcag gggtagaccc cagaatccat taagaaaa 2698
Claims (27)
1.一种汉尼巴病毒(Henipavirus)感染的金仓鼠动物模型,所述动物模型感染了汉尼巴病毒。
2.一种检测样品中的尼帕病毒(Nipah virus)的方法,其包括:
用至少一种特异于所述尼帕病毒的至少一种RNA分子的引物产生所述RNA分子的DNA拷贝;
用至少一对特异于所述尼帕病毒的RNA分子的所述DNA拷贝的寡核苷酸引物扩增所述DNA拷贝;和
检测对应于尼帕病毒的扩增的DNA的存在,所述扩增的DNA的存在表示样品中存在尼帕病毒。
3.权利要求2的方法,其中所产生并扩增的DNA拷贝是尼帕病毒核壳编码区。
4.权利要求2的方法,其中所述至少一对寡核苷酸引物包括一种可检测部分。
5.权利要求4的方法,其中检测包括观察所述可检测部分。
6.权利要求2的方法,其中样品取自猪。
7.权利要求2的方法,其中样品取自野生动物或家养动物。
8.权利要求2的方法,其中样品取自人。
9.权利要求2的方法,其中所述至少一种特异于所述RNA分子的引物包括与编码一种多肽的多核苷酸互补的至少15个连续核苷酸,所述多肽包括选自由SEQ ID NO:16、SEQ ID NO:23、SEQ ID NO:31和SEQ ID NO:32构成的组的氨基酸序列。
10.权利要求9的方法,其中所述至少一种特异于所述RNA分子的引物包括所述多核苷酸的至少20个连续核苷酸。
11.权利要求9的方法,其中所述至少一种特异于所述RNA分子的引物包括所述多核苷酸的至少25个连续核苷酸。
12.足以诱导免疫应答的量的至少一种分离的汉尼巴病毒(Henipavirus)G和F糖蛋白在制备用于保护个体或哺乳动物对抗汉尼巴病毒感染的药物中的用途。
13.权利要求12的用途,其中所述汉尼巴病毒是尼帕病毒(Nipahvirus)或亨德拉病毒(Hendra virus)。
14.权利要求12的用途,其中所述汉尼巴病毒G和F糖蛋白与一种佐剂结合。
15.一种表达载体在制备用于防护或保护个体或哺乳动物对抗汉尼巴病毒(Henipavirus)感染的药物中的用途,所述表达载体表达足以诱导免疫应答的量的至少一种分离的汉尼巴病毒G和F糖蛋白。
16.权利要求15的用途,其中表达载体至少表达汉尼巴病毒的F和G糖蛋白。
17.权利要求16的用途,其中表达载体是一种病毒载体。
18.权利要求17的用途,其中病毒载体是一种重组痘病毒载体。
19.权利要求15的用途,其中所述表达载体与至少一种佐剂结合。
20.一种产生抗汉尼巴病毒(Henipavirus)G或F蛋白之一或两者的抗体的重组杂交瘤。
21.一种表达汉尼巴病毒(Henipavirus)G或F蛋白之一或两者的重组痘病毒载体。
22.一种表达尼帕病毒(Nipah virus)G蛋白的重组痘苗病毒,其于2003年9月16日保藏于CNCM,保藏号为I-3086。
23.一种表达尼帕病毒(Nipah virus)F蛋白的重组痘苗病毒,其于2003年9月16日保藏于CNCM,保藏号为I-3085。
24.具有抗尼帕病毒(Nipah virus)的中和活性的抗尼帕病毒G蛋白的杂交瘤N°1.7,其于2004年9月9日保藏于CNCM,保藏号为I-3293。
25.具有抗尼帕病毒(Nipah virus)的中和活性的抗尼帕病毒G蛋白的杂交瘤N°3.B10,其于2004年9月9日保藏于CNCM,保藏号为I-3296。
26.具有抗尼帕病毒(Nipah virus)和亨德拉病毒(Hendra virus)的中和活性的抗尼帕病毒F蛋白的杂交瘤N°35,其于2004年9月9日保藏于CNCM,保藏号为I-3295。
27.具有抗尼帕病毒(Nipah virus)和亨德拉病毒(Hendra virus)的中和活性的抗尼帕病毒F蛋白的杂交瘤N°3,其于2004年9月9日保藏于CNCM,保藏号为I-3294。
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