KR20140043591A

KR20140043591A - 감자 유래 고온 스트레스 저항성 유전자 및 이를 이용하여 제조한 고온 스트레스 저항성 형질전환체

Info

Publication number: KR20140043591A
Application number: KR1020120108881A
Authority: KR
Inventors: 박세원; 유재웅; 사지쉬 카파처리; 히레마스 간가다르
Original assignee: 건국대학교 산학협력단
Priority date: 2012-09-28
Filing date: 2012-09-28
Publication date: 2014-04-10
Also published as: KR101604258B1

Abstract

본 발명은 감자 유래의 고온 스트레스 저항성 유전자 및 이를 이용하여 제조한 고온 스트레스 저항성 형질전환체에 관한 것으로서, 서열번호 1 내지 서열번호 95로 기재되는 유전자가 고온 스트레스 저항성을 갖는 감자에서 발현되는 것으로 확인됨으로써, 상기 유전자를 형질전환하여 고온 스트레스 저항성을 가지는 형질전환체를 제조할 수 있으므로, 상기 유전자는 고온에 대해 저항성을 획득한 식물체의 개발에 유용하게 이용할 수 있다.

Description

감자 유래 고온 스트레스 저항성 유전자 및 이를 이용하여 제조한 고온 스트레스 저항성 형질전환체{Heat stress resistant gene from potato and heat resistant transformant using the same}

본 발명은 감자로부터 유래하고 고온 스트레스에 저항성을 가지는 유전자 및 이를 이용하여 제조한 고온 저항성을 가지는 형질전환체에 관한 것이다.

감자(Solanum tuberosum)는 단위 경지 면적당 생산량이 옥수수, 밀, 쌀보다 더 많은 식량작물이며, 콜럼버스 이후 전세계에 전파되어 극지방을 제외한 대부분의 지역에서 재배되는 주요한 주식자원이다. 또한, 세계 감자 생산량의 반 이상이 개발도상국에서 생산되며, 이들 나라에서 감자는 음식과 농가소득에 중요한 비중을 차지하고 있다.

한편, 극한의 온도, 수분 부족, 염분의 과다, 미네랄 결핍 및 독성 등과 같이 환경적으로 가해진 스트레스에 의해 농작물의 생산성이 크게 저하되는 것으로 알려져 있다. 비생물적 환경 스트레스에 의한 농작물의 생산량 손실은 지대하여 가능한 생산량의 반 이상이 비생물적 스트레스에 의해 손실됨이 확인되고 있으며, 최근에는 국지적이나마 급격한 환경 변화가 기록되고 있어 이에 따른 작물의 피해 정도가 더욱 우려되고 있다. 이러한 환경적 스트레스에 의한 농작물의 피해를 줄이고 생산성을 향상시키기 위하여 일반적으로 농약이 사용되고 있으나, 농약의 사용은 오히려 환경 오염이라는 심각한 문제를 야기할 뿐만 아니라 이를 섭취할 경우 인체에도 해로운 영향을 준다. 그러나 환경적 스트레스에 의한 농산물 생산 감소 효과에 대해서는 식물에서 스트레스 내성의 근본적인 메커니즘과 관련하여 아직까지 명확하게 밝혀진 바가 거의 없는 실정이다. 감자에 있어서도, 생명공학 분야 연구는 매우 활발하여 역병 저항성 유전자연구, 마커를 이용한 유전자 지도 연구, 전분합성 변화에 의한 품질개선, 경구백신 생산 연구 등에서 괄목할만한 성과를 거두고 있으나 내재해성 연구는 아직 미진한 편이다(비특허문헌 1 참조).

환경적 스트레스에 궁극적으로 작용하는 식물체 세포 유전자 발현 기계로서 다양한 종류의 유전자들이 관여하고 있다. 이러한 식물내 환경적 스트레스에 대한 저항성 유전자를 발굴하기 위하여 지금까지의 연구에서는 돌연변이체 유도 및 탐색, 발현이 증폭되거나 억제되는 유전자에 대한 상호 비교 스크리닝(differential screening), 마이크로어레이, NGS를 이용한 전사체 비교 등의 방법에 의해 수행되어 왔으나, 최근에는 기존의 돌연변이 등과 같은 기능-상실(loss of function) 접근법 이외에 대장균, 효모 또는 아그로박테리아를 이용한 기능-획득(gain of function) 탐색법이 식물의 비생물적 스트레스 저항성 유전자의 탐색에 효과적임이 보고되고 있다(비특허문헌 2 참조).

그러나, 현재까지 감자를 대상으로 다양한 환경적 스트레스, 특히 고온 스트레스 저항성 유전자를 탐색한 연구는 수행되지 않고 있다.

Mullins et al., 2006 Eswaran et al., 2010 Hemavathi et al 2010 Burge and Karlin 1998 Salamov and Solovyev 2000 Altschul et al 1990

본 발명의 목적은 내열성이 뛰어난 유전자 및 이를 이용한 감자 품종을 육성하기 위하여, 감자 유래 고온 스트레스 저항성 유전자 및 이를 이용하여 제조한 고온 스트레스 저항성 감자 형질전환체를 제공하는데 있다.

이하, 본 발명에서 사용된 용어를 설명한다.

본 발명에서 사용된 용어 "고온 스트레스 저항성"은 높은 온도, 바람직하게는 35℃ 이상, 더욱 바람직하게는 39℃ 이상 45℃ 이하의 온도에서 생명체의 성장이 억제되지 않도록 하는 내성을 의미한다.

본 발명에서 사용된 용어 "형질전환체"는 유전공학적으로 조작된 것으로 외래 폴리뉴클레오티드 서열을 포함하며, 식물세포, 식물조직 또는 식물체에서 발현가능하도록 제작되어 외래 폴리뉴클레오티드 서열을 발현하는 것들을 의미한다.

이하, 본 발명을 상세히 설명한다.

상기 목적을 달성하기 위하여, 본 발명은 고온 스트레스 저항성 유전자를 포함하는 고온 스트레스 저항성 형질전환체 제조용 발현벡터로서, 상기 유전자는 서열번호 1 내지 서열번호 95로 이루어진 군으로부터 선택되는 유전자인 발현벡터를 제공한다.

상기 유전자는 서열번호 1 내지 서열번호 95 중 어느 하나의 염기서열 전체 또는 그것의 일부로 이루어질 수 있으며, 서열번호 1 내지 서열번호 95의 염기서열과 70% 이상, 바람직하게는 80% 이상, 더욱 바람직하게는 90% 이상, 가장 바람직하게는 95% 이상의 상동성을 가지는 서열일 수 있다. 상기 유전자는 식물체, 바람직하게는 감자(Solanum tuberosum)로부터 분리될 수 있다.

상기 발현벡터는 상기 유전자, 상기 유전자와 상동성을 가지는 유전자, 또는 다른 고온 스트레스 저항성 유전자를 추가로 포함할 수 있다. 상기 발현벡터로는 당업계에서 식물 형질전환용 발현벡터로 통용되는 것이라면 어느 것이든 사용 가능하다.

본 발명의 바람직한 구현예에 있어서, 상기 발현벡터는 프로모터 및 터미네이터를 포함할 수 있다. 상기 프로모터로는 식물 발현용 프로모터로서 CMV(Cauliflower Mosaic Virus) 35S 프로모터, CMV 19S 프로모터, 아그로박테리움 튜메팩시엔스 Ti 플라스미드(Agrobacterium tumefaciens Ti plasmid)의 nos(nopaline synthase) 프로모터, ocs(octopine synthase) 프로모터 및 mas(mannopine synthase) 프로모터 및 공지된 프로모터가 모두 사용될 수 있다.

본 발명의 다른 구현예에 있어서, 상기 발현벡터는 본 발명의 유전자의 발현을 확인하거나 형질전환체를 선별할 수 있는 선별 마커를 추가로 포함할 수 있다. 선별 마커로는 카나마이신, 히그로마이신, 젠타마이신 및 블레오마이신 등의 항생제에 대하여 내성을 나타내는 유전자, 또는 GUS(β-glucuronidase), CAT(chloramphenicol acetyltransferase), 루시퍼라제(luciferase) 또는 GFP(green fluorescent protein) 등을 암호화하는 유전자 등이 있으나 이에 한정되는 것은 아니다. 상기 선별 마커를 포함하는 발현벡터가 식물체에 전달되어 특정한 항생제를 포함하는 배지에서 배양함으로써 형질전환체의 선별을 가능하게 한다.

본 발명의 바람직한 구현예에 있어서, 본 발명자들은 고온 환경에 대한 적응성 후보 유전자를 발굴하고 이를 동정하여 이용함으로써 고온 스트레스에 저항성을 가지는 형질전환 식물체를 제조하기 위하여, 고온 스트레스 환경하에서 발현되는 감자 식물체 유래 유전자를 확인하였다. 이때, 효모 시스템을 이용한 cDNA 라이브러리의 기능-획득 스크리닝을 통해 고온 스트레스 저항성 유전자를 선발할 수 있다. 즉, 고온에 노출된 감자로부터 분리된 유전자의 cDNA를 효모 발현벡터에 클로닝하고(도 3 참조), 상기 발현벡터로 형질전환된 효모를 고온에 노출시킨 후, 고온 스트레스 내성 효모로부터 플라스미드를 분리하여 이로부터 고온 스트레스에 저항성을 갖는 유전자를 수득할 수 있다(도 4 및 도 6 참조). 상기와 같이 얻어진 유전자들을 서열 분석하여 동정한 결과, 하기 표 1 및 표 2와 같은 유전자들이 고온 스트레스 저항성을 갖는 효모내에서 발현되는 것으로 확인되었다(표 1 및 표 2 참조).

따라서, 본 발명의 서열번호 1 내지 서열번호 95로 기재되는 유전자가 감자에서 발현될 경우 고온 스트레스에 대한 저항성을 부여하는 유전자로서 유용하게 이용할 수 있다.

또한, 본 발명은 상기 발현벡터를 숙주 세포에 도입하여 제조한 형질전환체를 제공한다.

본 발명의 바람직한 구현예에 있어서, 상기 형질전환체는 식물체인 것이 바람직하며, 식물세포, 식물조직 또는 식물 종자를 포함한다. 상기 숙주 세포는 식물세포, 식물체 및 이들로부터 유래된 배양 세포 등을 포함한다.

상기 식물체는 단자엽 식물 또는 쌍자엽 식물일 수 있으며, 감자, 양파, 가지, 담배, 당근, 오이, 올리브, 고구마, 배추, 양배추, 무, 수박, 참외, 오이, 호박, 딸기, 대두, 녹두, 강낭콩, 상추, 고추, 토마토, 브로콜리, 해바라기, 갓, 잔디, 애기장대, 유채, 및 갓으로 이루어진 군으로부터 선택되는 식물인 것이 바람직하며, 감자인 것이 보다 바림직하나 이에 한정되지 않는다.

본 발명의 형질전환체는 공지된 기술로 제조할 수 있으며, 예를 들면 아그로박테리움(Agrobacterium sp.)-매개에 의한 방법, 유전자총(gene gun), 실리콘 탄화물 위스커법(Silicon carbide whiskers), 초음파 처리법(sonication), 전기천공법(electroporation) 또는 PEG(polyethylene glycol) 침전법 등의 방법으로 식물체에 본 발명의 유전자를 도입할 수 있다.

또한, 본 발명은, 상기 발현벡터를 제조하는 단계;

상기 발현벡터를 숙주 식물세포에 도입하여 형질전환 식물세포를 제조하는 단계; 및

상기 형질전환 식물세포를 조직 배양하여 재분화하는 단계를 포함하는 고온 스트레스 저항성을 갖는 식물체의 제조 방법을 제공한다.

상기 발현벡터를 숙주 식물세포에 도입하는 단계는 공지된 식물 형질전환 방법에 의하여 발현벡터를 식물체내로 도입할 수 있다. 즉, 아그로박테리움-매개에 의한 방법, 유전자총, 실리콘 탄화물 위스커법, 초음파 처리법, 전기천공법 및 PEG 침전법과 같이 식물체 형질전환할 수 있는 적합한 수단이거나 본 발명이 속하는 기술 분야의 기술자에게 알려진 공지의 방법이라면 어느 것이든 이용할 수 있다.

형질전환 후, 식물세포로부터 성숙한 식물로 재생될 수 있으며, 캘러스 또는 원형질체 배양으로부터 성숙한 식물의 재생을 위한 기술은 수많은 종에 대해 이미 본 기술분야에 잘 알려져 있다(Handbook of Plant Cell Culture, 1-5권, 1983-1989 Momillan, N.Y. 참조).

본 발명의 한 구현예에서, 본 발명자들은 식물체에 고온 스트레스 저항성을 증진시킬 수 있는 유전자를 얻기 위하여, 먼저 감자에 고온 스트레스를 가한 후, cDNA 라이브러리를 제작하여 고온 스트레스에서 발현되는 cDNA 클론들을 효모를 이용한 기능적 스크리닝 방법을 활용하여 선별한다.

따라서, 본 발명의 유전자를 숙주 세포에 도입하여 형질전환체를 제조할 수 있고, 이와 같이 제조된 형질전환체는 상기 유전자의 발현으로 인해 고온 스트레스 저항성을 가질 수 있다.

아울러, 본 발명은 상기 발현벡터를 식물체에 도입하는 단계를 포함하는 식물체의 고온 스트레스 저항성을 증가시키는 방법을 제공한다. 본 발명에 따른 식물체의 고온 스트레스 저항성을 증가시키는 방법은, 식물체의 고온 스트레스 저항성을 증가시키는 기능이 있는 것으로 확인된 감자 유래의 본 발명의 유전자들을 이용할 수 있다.

본 발명에 따르면, 감자로부터 분리한 본 발명의 유전자는 고온 스트레스에 의해 유도되는 유전자로서, 식물체 내에서 발현되도록 함으로써 식물체의 고온 스트레스 저항성을 증가시킬 수 있는 효과가 있다.

도 1은 감자 유래 고온 저항성 유전자를 스크리닝하기 위한 개략도이다.
도 2는 RNA 분리를 위하여 고온 스트레스(35℃) 하에서 다양한 시간 간격으로(30분, 2시간, 6시간, 12시간, 24시간 및 48시간) 회수한 감자 식물체를 나타내는 사진이다.
도 3은 본 발명에서 사용한 pYES2.1/V5-His-TOPO 벡터의 개략도이다.
도 4는 다양한 온도에 대한 효모의 생존성을 나타내는 사진이다.
도 5는 42℃ 고온 스트레스 하에서 본 발명의 형질전환체의 성장을 나타내는 사진이다.
도 6은 고온 스트레스 처리에 따른 유전자 발현량 변화를 나타내는 사진이다.

이하, 본 발명을 실시예에 의해 더욱 상세히 설명한다.

단, 하기 실시예는 본 발명을 예시하기 위한 것일 뿐, 본 발명의 내용이 하기 실시예에 의해 한정되는 것은 아니다.

< 실시예 1> 감자 식물체의 준비

감자(Solanum tuberosum) 품종 "태동밸리"를 2.5%(w/v) 수크로즈 및 0.8%(w/v) 식물용 한천(Duchefa, Germany)이 포함된 Murashige 및 Skoog(MS) 배지(Murashige and Skoog 1962)가 담긴 배양튜브에서 번식시켰다. 이 배양물을 22±2℃, 50~60%RH의 성장 챔버에서 백색 형광 램프에 의한 10 mmol/m²s 세기의 광을 16시간(명):8시간(암)의 광주기로 조사하여 유지하였다(비특허문헌 3 참조).

< 실시예 2> 고온 스트레스 처리

상기 실시예 1과 같이 시험관 내에서 성장한 3~4 주령의 어린 감자 식물체의 배지를 제거하고, 싹과 뿌리의 식물량을 기초로 하여 임의로 25~30개 소식물체씩 서로 다른 그룹으로 분리하였다. 이러한 식물체의 뿌리를 멸균수로 세척하고 비타민이 포함된 0.5×MS(Sigma-Aldrich) 및 0.5% 수크로즈 용액이 담긴 튜브에 옮겨 스트레스 개시 전까지 22±2℃, 60%RH의 성장 챔버에 배양하였다. 상기 소식물체를 신선한 배지에 옮긴 후, 35℃에서 48시간 이상 배양함으로써 고온 스트레스를 부여하였다. 그런 다음, 정해진 시점(30분, 2시간, 6시간, 12시간, 24시간 및 48시간 후)에 각 시료당 8~10개의 식물체를 회수하였다(도 2). 그 후 회수한 식물체를 액체 질소에 동결한 후 분석하기 전까지 -80℃에 보관하였다. 이때, 22±2℃에서 유지된 감자 식물체를 상기와 같은 시점에 각각 회수하여 대조군으로서 사용하였다.

< 실시예 3> cDNA 발현벡터의 제작

상기 실시예 2에서 준비한 각각의 감자 식물체로부터 효모 발현벡터를 이용하여 전장 cDNA 발현 라이브러리를 제작하였다.

<3-1> 총 RNA 의 분리

상기 실시예 2로부터 수득한 고온 처리한 식물 및 대조군 시료로부터 cDNA를 얻기 위하여, NucleoSpin RNA Plant mini prep Kit(Macherey-Nagel, Germany)를 사용하여 제조사의 지시에 따라 각각의 총 RNA를 분리하였다. 총 RNA의 순도는 포름알데히드 아가로스 겔을 통해 확인하였고, Nanodrop을 이용하여 측정하였다.

<3-2> cDNA 의 제작

상기 실시예 <3-1>에서 분리한 대조군 또는 고온 처리한 감자 식물체 유래 총 RNA로부터 Infusion SMARTer directional cDNA library construction kit(BD Clone Tech, USA)를 사용하여 방향성 cDNA 라이브러리를 제작하였다. 구체적으로, 제1가닥 cDNA는 제조사의 지침(Ref of Smart cDNA Kit)에 따라 3'Infusion SMARTer CDS 프라이머(12μM), SMARTer V 올리고뉴클레오티드(12μM), 5' PCR 프라이머Ⅱ A(12μM) 및 SMARTscribe 역전사중합효소(100 U/㎕)를 사용하여 폴리(A)⁺RNA 500 ng으로 합성하였다. 이중가닥 cDNA는 Accu power HF PCR Pre Mix(Bioneer, Korea)를 사용하여 긴 길이 PCR을 통해 합성하였고, 하기 프로그램에 따라 iCycler(Bio-Rad, USA)로 PCR 증폭을 실시하였다: 95℃에서 1분간 초기 변성; 95℃에서 15초간 변성 및 60℃에서 30초간 어닐링/연장 18 사이클, 및 68℃에서 6분간 최종 연장. 그런 다음, cDNA를 추출하였고, CHROMA SPIN + TE-1000 컬럼(BD Clone Tech, USA)을 사용하여 크기별로 분리하였다. 증폭 산물(amplicon)의 수율 및 순도를 1.2% 아가로스 겔 상에서 확인하였다.

<3-3> 효모 발현벡터의 제작

효모에서 고온 처리된 감자의 cDNA 라이브러리를 발현시키기 위하여, Amp^r에 대한 E. coli 선별 마커와 효모 선별 마커인 URA3 마커를 포함하는 E. coli-효모 셔틀 벡터인 효모 발현벡터 pYES2.1 TOPO TA(Invitrogen, Carlsbad, USA)를 사용하였다. In-Fusion SMARTer cDNA 라이브러리 제작을 위해 키트 프로토콜에 따라 효모 발현벡터 pYES2.1 TOPO TA를 선형화하였다. pYES2.1 TOPO TA 효모 벡터의 선형화는 Accu power HF PCR Pre Mix(Bioneer Corp, Korea)를 사용하여 프라이머 1; 5'-TTGATACCACTGCTTAGGGCGAGCTTAATATTCCCT-3'(10μM)(서열번호 96) 및 프라이머-2; 5'-TCTCATCGTACCCCGCTCCTCGGTCTCGATTCTACG-3'(10μM)(서열번호 97)를 포함하는 20 ㎕의 반응 부피에서 실시하였다. 그런 다음, 하기 PCR 프로그램에 따라 증폭을 실시하였다; 95℃에서 5분간 초기 변성; 95℃에서 1분간 변성 및 68℃에서 5.48분간 어닐링/연장을 20 사이클, 및 72℃에서 10분간 최종 연장. 증폭된 PCR 산물은 1.2% 아가로스 젤 상에서 분리한 후 Hiyield Gel/PCR DNA fragments extraction Kit (RBC, Real Genomics, Taiwan)의 프로토콜에 따라 pYES2.1 TOPO TA 플라스미드(5.8 Kb)의 겔 용출을 실시하였다.

<3-4> cDNA 발현 라이브러리의 제작

효모에서 라이브러리의 유전자 발현을 조절하기 위하여, Infusion HD cloning kit(BD Clone Tech, USA)를 사용하여 감자 식물체로부터 제작한 외부 cDNA를 선형화된 효모 벡터 pYES2.1 TOPO TA의 GAL1 프로모터의 다운스트림으로 클로닝하였다(도 3). pYES2.1 TOPO TA내 고온 처리한 감자의 cDNA 라이브러리를 E. coli HST08 premium Electro-cells(Takara Bio INC, Japan) 내로 형질전환하였다. 그런 다음, Infusion SMARTer directional cDNA library construction kit(BD Clone Tech, USA)에 제시된 바에 따라 재조합 클론의 비율을 확인하기 위하여 형질전환된 콜로니를 PCR을 사용하여 스크리닝하였다. 2시간 및 48시 동안 고온 처리한 감자 식물체로부터 각각 효모 GLA1 시스템하에서 조절되는 8.3×10⁶ 및 5.0×10⁶개의 개별적인 클론으로 구성된 2개의 cDNA 발현 라이브러리를 수득하였다. 그리고 나서, Gene All Exprep Plasmid Midi prep Kit(Gene All Biotech, Korea)를 사용하여 감자 식물체로부터 제작된 전체 라이브러리에서 플라스미드 DNA를 분리하였다.

< 실시예 4> 기능적 분석

<4-1> 효모 균주에 대한 기능적 스크리닝 온도의 결정

야생형 효모(Saccharomyces cerevisiae) 균주 BY4741(MATa; his3Δ1; leu2Δ0; met15Δ0; ura3Δ0)(EURO-SCARF, Germany)에 대한 허용적 또는 제한적 온도를 조사하기 위하여, 상기 효모를 합성 선별 배지(D-갈락토스 20 g/L , 라피노스 0.5 g/L , 우라실이 없는 효모 합성 드롭아웃 배지(Sigma Aldrich) 1.92 g/L, 황산암모늄이 포함된 효모 니트로겐 베이스(MP Biomedicals, France) 6.7 g/L)에 30℃에서 하룻밤 동안 배양하였다. 10^-3으로 희석한 상기 배양물 100 ㎕를 2% 한천이 포함된 합성 선별 배지에 도말하여 30 내지 39℃ 범위의 서로 다른 온도에서 배양하였다.

그 결과, 39℃에서 배양한 효모 균주의 경우 95% 이상에서 성장 결함이 관찰되어, 39℃를 효모의 스크리닝 온도로서 선택하였다(도 4).

<4-2> 효모로의 라이브러리 플라스미드의 형질전환 및 고온 스트레스의 처리

효모 GAL1 프로모터에 의해 조절되고, 감자 식물체 유래의 클로닝된 삽입체(inserts)를 포함하는 효모(pYES2.1 TOPO TA) 플라스미드를 야생형 효모 균주 BY4741에 Biorad Micro Pulser(Biorad, USA)를 사용하여 전기천공법을 통해 형질전환하였다(L. Benatuil et al 2010 참조). 전기천공 후 즉시, 상기 효모를 1M 소르비톨(Sigma Aldrich)과 YPD 배지가 1:1로 혼합된 혼합물 8 ㎖에 옮기고, 225 rpm으로 1시간 동안 30℃의 진탕배양기(shaking incubator)에서 배양하였다. 최종적으로 3000 rpm으로 3분 동안 원심분리를 하여 세포를 수득하고 펠릿을 250 ㎕의 합성 선별 배지에 재현탁하였다. 그 결과 얻어진 세포 혼합액을 2% 마이크로 한천(Duchefa, Netherlands)이 포함된 합성 선별 배지에 도말하여 고온 스트레스를 주기 위해 3~4일 동안 39℃에서 배양하였다. 대조군으로서 효모 형질전환체를 포함하는 혼합액을 상기 배지에 도말하여 3~4일 동안 30℃에서 배양하였다.

<4-3> 효모 기능적 스크리닝

고열 처리한 감자의 cDNA를 발현하는 효모 형질전환체는 야생형 효모 균주 BY4741 하에서 39℃의 온도(효모 BY4741 성장에 제한적인 온도)에서 배양하여 분리하였고, 음성 대조군(pYES2.1 TOPO TA 빈 벡터)과 비교하여 고온 저항성을 가지는지 확인하기 위하여 동일한 배지에 도말하였다.

그 결과, 이종 감자 cDNA를 발현하는 URA3 마커를 갖는 플라스미드에 의해 약 48,000개 이상의 효모 형질전환체가 성장한 것을 확인하였다. 2시간 및 48시간 동안 각각 고온 처리한 감자 유래 cDNA를 발현하는 120 및 150개의 각 후보 효모 형질전환체(39℃에서 성장하는 고온 저항성 클론)를 수득하였다. 상기 각각의 개별 효모 형질전환체의 고온 저항성을 더 확인하기 위하여, 이를 합성 선별 배지가 포함된 플레이트상에 도말하여 39℃ 및 42℃에서 추가 배양하여 상기 형질전환체가 생존하는 것을 확인하였다(도 5).

<4-4> 플라스미드 분리 및 E. coli 로의 역형질전환

플라스미드 DNA를 선행문헌에 보고된 방법(Byrd and St-Arnaud, 2001)을 약간 수정하여 pYES2.1 TOPO TA 효모 벡터에 삽입체를 포함하는 상기 고온 저항성 효모 형질전환체로부터 분리하였다. 구체적으로, 상기 고온 저항성 효모 형질전환체를 포화될 때까지 2~3일 동안 덱스트로스를 포함하는 합성 선별 배지에 30℃ 온도에서 배양하였다. 그런 다음, 상기 형질전환체를 0.3 g의 산 세척된 유리 비드, 효모 용해 용액(2%(v/v) 트리톤 X-100, 1%(w/v) 도데실황산나트륨, 100 mM NaCl, 10 mM Tris-HCL(pH 8.0), 1 mM EDTA) 200 ㎕, 및 클로로팬(페놀:클로로포름:이소아밀알코올=25:24:1) 200 ㎕을 첨가하여 5분 동안 볼텍싱(vortexing)하여 용해하였다. 클로로팬 추출, 이어서 핵산의 에탄올 침전 플라스미드을 1회 이상 실시하여 상기 효모로부터 플라스미드 DNA를 분리하였다.

pYES2.1 TOPO TA 효모 벡터에서 클로닝된 상기 삽입체를 더 분석하기 위하여, 고온 저항성 효모 형질전환체로부터 분리한 상기 플라스미드 DNA를 전기천공법을 통해 E. coli로 다시 형질전환하였다.

< 실시예 5> DNA 서열분석 및 주석( annotation )

Big Dye^TM Terminator Cycle Sequencing Kit(ABI, USA) 및 DNA analyzer(ABI 3730XL, USA)를 사용하여 이종 cDNA를 발현하는 고온 저항성 플라스미드로부터 분리한 cDNA의 서열을 분석하였다. 시퀀싱 반응은 GAL1 정방향 시퀀싱 프라이머 5'-AATATACCTCTATACTTTAACGTC-3'(서열번호 98)을 사용하여 실시하였고, 추가 분석 전에 VecScreen (http://www.ncbi.nlm.nih.gov/VecScreen/VecScreen.html)을 사용하여 벡터 및 어댑터 서열을 잘라냈다. 전처리 후, GENSCAN (비특허문헌 4 및 비특허분헌 5 참조)을 통해 가능성 있는 유전자로 확인되지 않는 서열들에 대하여 GeneScan(http://genes.mit.edu/GENSCAN.html) 및 FGENESH(http://linux1.softberry.com/berry.phtml?topic=fgenesh&group=programs&subgroup=gfind)를 사용하여 가능성 있는 전사해독틀(open reading frames)을 예측하였다. 또한, 유전자 기능을 확인하기 위하여, BLAST 알고리즘을 사용하여 NCBI (http://www.ncbi.nlm.nih.gov/blast/) 및 TIGR plantt ranscript assemblies (http://tigrblast.tigr.org/euk-blast/plantta-blast.cgi)에 있는 서열 데이터베이스에 대한 상동성 검색을 실시하였다(비특허문헌 6 참조).

그 결과, 고온 저항성 플라스미드로부터 분리한 cDNA 서열은 하기 표 1 및 표 2에 기재된 유전자와 매칭되는 것으로 확인됨으로써, 하기 유전자들은 감자 식물체에서 고온 스트레스 저항성에 관여하는 유전자인 것을 알 수 있었다.

2시간 고온 스트레스를 처리한 고온 저항성을 갖는 감자 식물체로부터 효모 기능적 스크리닝을 통해 획득한 cDNA 서열

서열번호	클론 ID	등록번호 (Accession number)	추정상 유전자 기능	유전자 길이 (bp)	전장*
1	St-TNT1	JX576196	14 kDa proline-rich protein DC2.15	402	Y
2	St-TNT2	JX576197	ribosomal protein L36	327	Y
3	St-TNT6	JX576198	aquaporin (PIP3)	822	Y
4	St-TNT8	JX576199	putative chaperon P13.9	399	Y
5	St-TNT9	JX576200	fiber protein Fb15	264	Y
6	St-TNT11	JX576201	60S ribosomal protein L37a	278	Y
7	St-TNT13	JX576202	60S ribosomal protein L13	621	Y
8	St-TNT14	JX576203	60S ribosomal protein L33	339	Y
9	St-TNT15	JX576204	light-inducible protein ATLS1	348	Y
10	St-TNT16	JX576205	light inducible tissue-specific ST-LS1 gene	417	Y
11	St-TNT20	JX576206	hydroxyacylglutathione hydrolase	774	Y
12	St-TNT22	JX576207	putative proteinase inhibitor type II (Pi7Ca)	458	Y
13	St-TNT23	JX576208	chloroplast photosystem I reaction center V gene	440	Y
14	St-TNT24	JX576209	chlorophyll a-b binding protein 3C-like mRNA	702	Y
15	St-TNT31	JX576210	Cab-4 gene encoding chlorophyll a/b-binding protein	816	Y
16	St-TNT36	JX576211	Ribosomal protein L24	495	Y
17	St-TNT41	JX576212	putative Rieske Fe-S protein precursor (FeS) gene	693	Y
18	St-TNT42	JX576213	chloroplast manganese stabilizing protein-II (Msp-II)	882	Y
19	St-TNT46	JX576214	pathogenesis-related protein P2	432	Y
20	St-TNT48	JX576215	ultraviolet-B-repressible protein	360	Y
21	St-TNT51	JX576216	Ribosomal protein L32	399	Y
22	St-TNT54	JX576217	plastocyanin precursor (pete gene)	515	Y
23	St-TNT55	JX576218	LOB domain protein 40	734	Y
24	St-TNT58	JX576219	chloroplast genes for maturase, trnK for tRNA-Lys(UUU), rps16 for ribosomal protein S16	546	Y
25	St-TNT64	JX576220	photosystem I subunit XI mRNA	642	Y
26	St-TNT69	JX576221	Hypothetical protein	318	Y
27	St-TNT71	JX576222	putative chloroplast protein CP12	369	Y
28	St-TNT78	JX576223	Solanum tuberosum Conserved gene of unknown function (PGSC0003DMT400047309)	291	Y
29	St-TNT83	JX576224	glutathione S-transferase-like protein	681	Y
30	St-TNT85	JX576225	Putative iojap protein	702	Y
31	St-TNT88	JX576226	ubiquitin-conjugating enzyme	446	Y
32	St-TNT30	JX576227	60S ribosomal protein L4/L1	888/1220	N
33	St-TNT34	JX576228	putative non-specific lipid transfer protein	218/335	N
34	St-TNT39	JX576229	putative spermidine hydroxycinnamoyl transferase	1160/1320	N
35	St-TNT40	JX576230	fructokinase	771/990	N
36	St-TNT47	JX576231	phototropin-1 (PHOT1)	222/3057	N
37	St-TNT59	JX576232	putative rac GTPase activating protein	180/1536	N
38	St-TNT68	JX576233	neutral invertase like protein, mRNA	600/1380	N
39	St-TNT81	JX576234	cold-stress inducible protein (C17)	651/1713	N
40	St-TNT90	JX576235	NAD(P)H-quinone oxidoreductase subunit 4	275/1143	N

48시간 고온 스트레스를 처리한 고온 저항성을 갖는 감자 식물체로부터 효모 기능적 스크리닝을 통해 획득한 cDNA 서열

서열번호	클론 ID	등록번호 (Accession number)	추정상 유전자 기능	유전자 길이 (bp)	전장
41	St-DNT3	JX576236	Germin-like protein subfamily 1 member 20 precursor	687	Y
42	St-DNT8	JX576237	putative non-specific lipid transfer protein	345	Y
43	St-DNT16	JX576238	chlorophyll a/b binding protein (Lhcb1-2) gene	839	Y
44	St-DNT17	JX576239	17.6 kD class I small heat shock protein	465	Y
45	St-DNT24	JX576240	40S ribosomal protein S9	594	Y
46	St-DNT31	JX576241	Myo-Inositol oxygenase	909	Y
47	St-DNT32	JX576242	E3 ubiquitin ligase (RING1)gene	557	Y
48	St-DNT35	JX576243	photosystem I subunit III precursor (psaF gene)	687	Y
49	St-DNT36	JX576244	TAS14 dehydrin	438	Y
50	St-DNT45	JX576245	RNase H family protein	591	Y
51	St-DNT46	JX576246	calmodulin	449	Y
52	St-DNT47	JX576247	40S ribosomal protein S5	660	Y
53	St-DNT48	JX576248	autophagy 8h gene	360	Y
54	St-DNT50	JX576249	Avr9/Cf-9 rapidly elicited protein 65	285	Y
55	St-DNT51	JX576250	light inducible tissue-specific ST-LS1 gene	417	Y
56	St-DNT53	JX576251	eukaryotic initiation factor 5A3	483	Y
57	St-DNT58	JX576252	Beta-fructofuranosidase	897	Y
58	St-DNT64	JX576253	Basic helix-loop-helix protein BHLH3	912	Y
59	St-DNT70	JX576254	Thylakoid lumenal 17.4 kDa putative protein	697	Y
60	St-DNT71	JX576255	ethylene-responsive element binding protein	693	Y
61	St-DNT74	JX576256	FK506-binding protein (FKBP)	450	Y
62	St-DNT76	JX576257	glutamine synthetase GS1 (gln)	1032	Y
63	St-DNT77	JX576258	wound-induced gene WIN1	636	Y
64	St-DNT81	JX576259	Stress-associated protein 3 (SAP3) gene	515	Y
65	St-DNT93	JX576260	Ribosomal protein L1	690	Y
66	St-DNT95	JX576261	60S ribosomal protein L35	363	Y
67	St-DNT96	JX576262	Potato Transmembrane and coiled-coil domains protein	585	Y
68	St-DNT97	JX576263	ribosomal protein L27	408	Y
69	St-DNT107	JX576264	RNA binding protein	654	Y
70	St-DNT109	JX576265	protease inhibitor-related protein (PI)	237	Y
71	St-DNT113	JX576266	auxin-repressed protein (ARP1)	384	Y
72	St-DNT121	JX576267	stress responsive cyclophilin protein	516	Y
73	St-DNT123	JX576268	chloroplast photosystem I reaction center V	441	Y
74	St-DNT142	JX576269	Auxin repressed/dormancy associated protein	375	Y
75	St-DNT9	JX576270	Geranylgeranyl reductase	744/1395	N
76	St-DNT25	JX576271	consereved gene of unknow function	747/1170	N
77	St-DNT26	JX576272	UDP-glucose:protein transglucosylase (uptg1 gene)	510/1087	N
78	St-DNT27	JX576273	Ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit (rbcL) gene	579/1080	N
79	St-DNT28	JX576274	Solanum tuberosum unknown Transcription factor	600/1221	N
80	St-DNT34	JX576275	Phosphoenolpyruvate carboxylase	870/2874	N
81	St-DNT62	JX576276	phosphoinositide-specific phospholipase C	792/1485	N
82	St-DNT79	JX576277	HAD superfamily protein involved in N-acetyl-glucosamine catabolism	671/957	N
83	St-DNT83	JX576278	ATP phosphoribosyl transferase	828/1185	N
84	St-DNT86	JX576279	fructokinase	801/990	N
85	St-DNT87	JX576280	conserved gene of unknown function	884/978	N
86	St-DNT88	JX576281	TAF-3 protien	648/1014	N
87	St-DNT92	JX576282	elongation factor 1-alpha-like protein	930/1347	N
88	St-DNT94	JX576283	Hydrolase, hydrolyzing O-glycosyl	598/2595	N
89	St-DNT102	JX576284	ASR4	650/894	N
90	St-DNT104	JX576285	40S ribosomal protein S2	768/831	N
91	St-DNT105	JX576286	Expansin-like B1 precursor protein	399/753	N
92	St-DNT117	JX576287	CDSP32 protein (Chloroplast Drought-induced Stress Protein of 32kDa)	549/891	N
93	St-DNT124	JX576288	conserved gene for unknown function	471/1656	N
94	St-DNT134	JX576289	DNAJ-like protein	269/467	N
95	St-DNT146	JX576290	Oxidoreductase	708/1086	N

< 실시예 6> qRT - PCR 을 통한 전사체 분석

NucleoSpin RNA Plant mini prep Kit(Macherey-Nagel, Germany)을 사용하여 서로 다른 시간 간격(0시간, 2시간, 24시간 및 48시간)으로 고온(35℃)을 처리한 총 RNA를 3~4주 자란 감자 식물체로부터 추출하였다. cDNA 제작을 위하여, ccuPower RocketScript RT PreMix Kit (Bioneer, Korea)에 따라 RocketScript 역전사중합효소(Bioneer, Korea) 및 oligoDT를 사용하여 약 750 ng의 총 RNA로부터 제1가닥을 합성하였다. RT-PCR은 5 ㎕ cDNA 주형 및 1 ㎕의 유전자 특이적 프라이머를 포함하는 Accupower® 2x greenstar qpcr master mix를 사용하여 실시하였다. 상기 RT-PCR은 하기 조건에 따라 Bio-Rad CFX96 sequence detector(Bio-Rad, USA)를 사용하여 실시하였다; 95℃에서 10분; 95℃에서 20초, 60℃에서 45초를 29 사이클. 이때, 감자 액틴 유전자를 유전자 발현 분석을 위한 내부 표준으로서 사용하였다. 서로 다른 유전자에 대한 총 5개의 시료를 RT-PCR 분석을 실시하였다. 그 결과 수득한 상대적 한계값(comparative threshold, Ct)을 상기 액틴에 대하여 표준화하였고 상이한 시간 간격에 수득한 상기 시료 유래의 서로 다른 유전자의 상대적 발현량을 계산하였다.

그 결과, plastocyanin precursor(pete gene), chaperon P13.9, LOB domain protein 40, hydroxyacyl glutathione hydrolase(HAGHL), proteinase inhibitor type II(Pi7Ca), 60S ribosomal protein L13, chlorophyll a-b binding protein, chloroplast manganese stabilizing protein-II(MSP II), TAS14 dehydrin, 40S ribosomal protein S9, E3 ubiquitin ligase(RING1), photosystem I subunit III precursor (psaF gene), glutamine synthetase GS1(gln) 및 Calmodulin protein 유전자의 감자 식물체 내 발현량은 고온 스트레스 처리 여부 및 처리 시간에 따라 변화하는 것으로 나타났고, 특히 고온 스트레스 노출시 발현이 현저하게 증가하는 유전자들을 확인하였다(도 6).

<110> Konkuk University Industrial Cooperation Corp <120> Heat stress resistant gene from potato and heat resistant transformant using the same <130> 2012-DPA-264 <160> 98 <170> KopatentIn 2.0 <210> 1 <211> 402 <212> DNA <213> Solanum tuberosum <400> 1 atggcttcca aacacatttt cccttgtctt gctattttcc tttgctttaa ccttttcttt 60 gccttagttt ctagttgtgg cacttgtcct aaacctaaac caaaaccaaa gccatcatgc 120 ccaccaccac cttattatcc taaagaaact tgtccaattg atacacttaa gttaggtgtt 180 tgtgctgatg ttcttggatt agtaaatgtt attgttggct caccaccagt aactccatgt 240 tgtagtcttc tttctggact tgctaatgct gaagctgcta tttgtctttg tactgctctt 300 aaggctaata ttttggggat taaccttaat ctccctattt cacttagttt gcttcttaat 360 gtttgctcta aagaagctcc tgctggcttc caatgctctt aa 402 <210> 2 <211> 327 <212> DNA <213> Solanum tuberosum <400> 2 atggcgccgc cgaacacagg attagccgtg ggattgaaca aaggacacat tgtaaccaag 60 aaggagctag ctccacgccc ttctgacaga aaagggaaaa caagcaaaag aatccacttc 120 gttaggaacc tcatcagaga agttgctgga tttgccccat atgagaagag gattactgag 180 cttcttaaag ttggcaagga caagcgtgca ttgaaggttg ccaagaggaa gttgggtacc 240 cacaagaggg caaagaagaa gagagaggag atgtctagtg ttctccgtaa gatgagggcc 300 accggaggtg gtgaaaagaa gaagtga 327 <210> 3 <211> 678 <212> DNA <213> Solanum tuberosum <400> 3 atggtgaaaa actatgttga tccaccttca gctccgctgt ttgatacagc ggagctgaac 60 aactggtctt tttacagagc attgattgct gaatttgttg cgacgcttct tttcctctat 120 gttagcgtgg ctactgttat tggccacaag aagcaagttg gtccctgcga cggcgttgga 180 cttgttggta ttgcatgggc ttttggtggc atgatttttg ttcttgtcta ttccactgct 240 gggatttctg gtgggcatat aaacccagca gtaacgtttg ggctattact agcaaggaag 300 gtgtcattac taacagcggt ggcgtacatg gtggcgcaat gcttaggagc tatttgcggc 360 gttggtttag tgaaaagtgt gatgaaggat gactataaca cgcatggtgg tggtgctaat 420 accgtcgcag ttggctactc aaccggcgca gcattaggtg ctgagattat tggcactttc 480 cttctcatgt ataccgtatt ctccgccaca gacgccaaaa gaaaagcacg tgattcccac 540 gtccctgtac tagcaccatt gccaattgga ttctcggtgt ttatggttca cttggccacc 600 attcccatca ctggcactgg catcaatcct gctaggagtt tcggagctgc tgtcatttac 660 aacgatacca cagcttga 678 <210> 4 <211> 399 <212> DNA <213> Solanum tuberosum <400> 4 atggcaaatt ctgtgtgttt caccccttta gcttcattga acactttcaa taaaccaggt 60 ttaattaatg gaaatgggaa ttgcgctgga agaaagattc aattgatcaa agatgttacc 120 ttcaactcca agagtaactt aagagtcgta gaagtgaagg ccgctgacag tgataaagaa 180 acaaaagtaa ggagcatagt ctgtcaaaag tgtgagggaa atggtgcagt ggcgtgctca 240 caatgtaaag gcgttggggt taactccgtc gatcatttca acggacggtt taaagcaggt 300 ggactatgtt ggctgtgcag aggtaaaaaa gatatattgt gtggtgactg caatggagct 360 ggatttcttg gtggatttat gagtacgttc gatgagtag 399 <210> 5 <211> 264 <212> DNA <213> Solanum tuberosum <400> 5 atggctttga gaaatttgta caaggagatg aagggattga agctgaagga ggtgccggcg 60 cacctaaagc cgatgctctc aatcggttac gcgaagaatg ctatcaggaa gggtttggat 120 aactaccatg ctaagtacat cgaaaccagc tccgttgatc ctcttctcca tgtctgcttc 180 ggaggcatga tattctccta cctcgtcgcc cttccagaag agcgacgtca cctcgagcac 240 aaacagcacg gcggcggaca ttga 264 <210> 6 <211> 279 <212> DNA <213> Solanum tuberosum <400> 6 atgactaaga ggaccaagaa ggctggaata gttggggaat atggtacccg gtatggtgcc 60 agtctgcgaa agcagattaa gaaaatggag gttagccagc atagcaagta cttctgcgag 120 ttttgtggaa agtatgcagt gaagaggaaa gccgtgggta tttggggatg taaagattgc 180 ggcaaagtca aagcaggcgg tgcttataca ttgaacactg caagtgctgt gacagttagg 240 agcacaatcc gaagactgag ggagcaaact gagagttag 279 <210> 7 <211> 621 <212> DNA <213> Solanum tuberosum <400> 7 atgaagcaca acaatgttat tcccaatggg cacttcaaga agcattggca gaactatgta 60 aggacatggt tcaaccaacc tgctaggaaa acaaggagac gtgctgctag gcagcagaag 120 gctgtgaaga tcttccctag gccaacagct ggttcacttc gacctattgt tcatggacaa 180 acccttaaat ataacatgaa agtccgtgct ggcaggggat tctctcttga agaactgaag 240 gctgctggta ttcccaaaaa gctagcacca accataggca ttgctgttga tcatcgccgc 300 aggaacagat ccttggaagg tctccaaacc aatgtccaga ggcttaagac ctacaaggct 360 aagctagtta tcttcccaag gcgtgctaag aaggtcaagg ctggtgattc tagtgcagag 420 gaacttgcta ctgccaccca ggttcagggt tcatacctgc ctattactag ggagcagcct 480 gctgttgact ttgtgaaggt cacagatgag atgaaatcat tcaaggccta tggcaagctt 540 cgtattgagc gtacaaatgc tcgacatatg ggagccaggt tgaagagggc agctgaagca 600 gaaaaggaag agaagaaata a 621 <210> 8 <211> 339 <212> DNA <213> Solanum tuberosum <400> 8 atggtgaagg gacgccaagg agagcgtgta agactctata ctagaggaac agtccttgga 60 tacaagaggt cgaagtcgaa tcagtatcca aacacttcat tggtgcagat cgagggagtg 120 aacactaagg aagaagtaga ttggtacttg gggaagcgta tggcttatgt atacaaggcg 180 aaaacaaaga agaaaaactc gcattaccgc tgcatttggg gtaaagtttg taggcctcat 240 ggtaacagtg gtgtcgttag agctaaattc gagtccaatc tgccaccaaa atccatggga 300 gctaaggtta gagttttcat gtacccaagc aatatataa 339 <210> 9 <211> 348 <212> DNA <213> Solanum tuberosum <400> 9 atgccgtgcc ttaacatttc tacaaacgtg aacttggaag gagttgatac ttcctccgta 60 ctttctgaag ctacctccac tgttgccaaa ctcatcggca aaccagaagc ttatgtcatg 120 attgtgttga agggatctgt tcccatggct ttcgggggta cagaacagcc cgctgcctat 180 ggagagttgg tctccattgg gggattaaat gctgatgtca ataagaaact cagtgctgca 240 atagctgaca tacttgagac taaattgtct atccccaaat ctcgattttt cctgaaattc 300 tatgatgcta agggttcctt ctttggctgg aatggatcta ccctctga 348 <210> 10 <211> 417 <212> DNA <213> Solanum tuberosum <400> 10 atggcaagca cagtaatgag ctcattgagc ctaaagccaa ctttcacttt ggagaaaaca 60 tcagtgaaag ggcttccatc acttgctaga tcttcttctt ccttcaaagt tgtggctagt 120 ggtgtcaaga agcttaagac tgacaaaccc tatggaatta atggaagcat ggccttgaga 180 gatggcgttg atgcctcagg caggaagccc aagggaaagg gtgtgtacca atatgttgac 240 aaatatggag ctaatgttga tggatacagt cccatctaca acacagatga gtggtctcca 300 agtggtgatg tctatgttgg aggtactact ggcttagcca tatgggccgt gaccttggtt 360 ggtattcttg caggaggtgc tctccttgtc tacaacacaa gtgctttggc acagtag 417 <210> 11 <211> 774 <212> DNA <213> Solanum tuberosum <400> 11 atgaaagttc ttcacatccc atacttagat gacaactact cctatatgat catcgatgaa 60 actactaagg aagctgcggt ggtagaccct gtcgagccgc acaaggtcct cagagttgct 120 caagagaacg aagttgacct caagtttgtt ctcaccactc atcaccactg ggatcatgct 180 ggaggaaatg ataaaataaa gcagctcgtt cctggaataa aaatttatgg tggatcagtt 240 gataatgtga ggggttgtac agataaagtt gaaaatggtg acagggtatc ccttggtgtt 300 gacatcagca tcttatctct tcacacacca gcccacacta aaggtcatat aagctactat 360 gttactgaca aagagggaga ggatcctgct gtttttactg gggatacatt gtttatagct 420 ggttgcggta agttttttga aggcactgca gaacaaatgt atcaatcact ctgtgtgaca 480 ttaggttctt tgccaaagcc aactctggta tattgtggtc atgagtacac agtaaaaaat 540 ttgcagtttg cattaacagt tgaaccggac aatgccaaaa ttgcacaaaa gttggcatgg 600 gctgagcgtc agagaagagt aggccttcca actattccat cgactattca ggaagaattt 660 gagagtaacc cattcatgcg ggttgatctt ccagaggttc agataaagtg gggtgcaagt 720 ctcctgttga agccatgcgt gaaatcagca acgaaagata attggaaggg ctga 774 <210> 12 <211> 459 <212> DNA <213> Solanum tuberosum <400> 12 atggctattc acaaagttgc tctcctcctt atatttggaa taattcttct tgcaagcgat 60 ttcgaacatg cagacgccaa agaatgtgac cctcgaattg actttggaat ttgcccatat 120 ttaggaacca aaaaagttga tggaatatgc atcaattgtt gttcgggtta caaagaatgt 180 aaatatttta gtaaagatta cacttttatt tgtgatggag aatctacaga ttggcaaagc 240 gggaaggctt gctcaaaaga atgtgaccgt cgaattgagt ttggaatttg cgtaacagac 300 ctaactaaaa ctgttaatgg actatgcacc acctgttctg caggtaaaaa aggatgtaaa 360 tatttcaata acaatggaac ttatatttgt gatggagaat atgaatggct aagtgaagga 420 gaaaatgatc tccagaaaag caatgttgct atttcttaa 459 <210> 13 <211> 441 <212> DNA <213> Solanum tuberosum <400> 13 atggcatcag ctctgttttc tgcaccaact ttccaaggtc tcagaccact caacaaaccc 60 actgactctt ccctcttcct caacaaaccc atcgccttca aacctatcac caaaaaaacc 120 cgaaatctcg ctgtgaaagc tgaactcaac ccatcaattg tcatcagttt gagcactggg 180 ctttcgcttt tccttggaag gttcgttttc tttaatttcc agagagaaaa tgtagcgaaa 240 caagtgccgt ctcagaatgg aatttctcac tttgaagctg gagatgttag ggctaaggaa 300 tatataagtt tgctaaaatc taatgatcca gttggattca atattgttga tgtgcttgct 360 tggggatcta ttggacatat tgttgcttat tatatccttg ctacttccag caatggatat 420 gatcccagtt tctttggctg a 441 <210> 14 <211> 702 <212> DNA <213> Solanum tuberosum <400> 14 atgaggaaga ctgctgccaa ggcaaagcct gcctcctctg gtagcccatg gtacggtcct 60 gaccgtgtta agtacttggg cccattctct ggtgaatccc caagctactt gactggtgag 120 ttccctggtg actacggatg ggacactgct ggactttcag ctgaccctga aacattcgct 180 aagaaccgtg aattggaggt gattcactgc agatgggcca tgcttggagc tcttggatgc 240 gtcttccctg agcttttggc ccgtaacggt gtcaagtttg gtgaagctgt atggttcaag 300 gctggatccc aaattttcag cgagggtgga ctcgactact tgggaaaccc aagcttggtc 360 catgcacaaa gcatcttggc tatttgggct tgccaagttg tgttgatggg agccgttgaa 420 ggttaccgta ttgctggtgg gcctcttggt gaggttgttg acccacttta ccctggtggt 480 agctttgacc cattgggcct tgctgacgac ccagaggctt ttgctgagct caaggttaag 540 gaaatcaaga atggcagact tgctatgttc tccatgtttg gattctttgt ccaagctatc 600 gtcaccggaa agggtccatt ggagaacctt gctgaccacc ttgctgaccc agttaacaac 660 aacgcctggg cctttgccac aaacttcgtt cccggaaaat ga 702 <210> 15 <211> 816 <212> DNA <213> Solanum tuberosum <400> 15 atggccactt ctgcaattca acaatcagcc tttgttggtc aagcagttgg taaatctcaa 60 aatgagtttg ttaggaaagt tggcagctgc ggtgaaagcc gtattaccat gaggcgtact 120 gttaaaagcg ctcctcaaag catctggtac ggagaagacc gtcctaagta cttgggccca 180 ttttctgagc aaacaccatc atactcgact ggtgaattcc caggtgacta tggatgggac 240 actgctggac tttcagctga cccggagaca tttgctagaa accgtgaact tgaggtgatc 300 cactgccgtt gggctatgct tggtgctttg ggctgtgttt tccccgaaat tctttcaaag 360 aacggtgtta aattcggtga ggctgtttgg ttcaaggctg gttctcagat tttctcagaa 420 ggtgggcttg actaccttgg aaacccaaac cttgtccatg cccaaagcat tcttgccatt 480 tgggcctgcc aagttgtcct tatgggcttt gttgaaggat acagagttgg tggaggccca 540 cttggtgagg gtcttgacaa gatctaccca ggaggtgcct ttgacccact cggtcttgct 600 gatgatcccg aggcttttgc tgaattgaag gtcaaggaaa ttaagaacgg acgtttggct 660 atgttctcca tgtttggatt ctttgttcaa gctattgtta ctggaaaggg cccaattgag 720 aacctttctg accacattaa tgacccagtg gctaacaacg cttgggctta cgccacaaac 780 tttgtacctg aaagtgaagt aattaataga atgtaa 816 <210> 16 <211> 495 <212> DNA <213> Solanum tuberosum <400> 16 atggttctca agacagagct ttgccgtttc agtggtgcca agatttaccc tgggagaggc 60 atcagattta tccgttcaga ttctcaggtg ttcctattcg tcaactcaaa gtgcaaacac 120 tacttccaca acaagttgaa gccgtccaac cttacatgga cagctatgta taggaagcaa 180 cacaagaagg atattgcaca agaagctgct aagaagaggc gacgtaccac caagaagcct 240 tactctaggt ctattgtggg tgcaaccttg gaggttatcc agaagaagag aactgaaagg 300 ccagaagttc gagatgctgc cagggaggct gctcttcgtg aaattaagga aaggatcaag 360 aagacaaagg atgaaaagaa ggcgaagaaa gcagaggttc aggccaagtc ccagaaagct 420 ggaggaaagg gcaatatgtc caaaggaggt gcatctaaag gtcctaagct tggtggtggc 480 ggaggaaaac gttaa 495 <210> 17 <211> 693 <212> DNA <213> Solanum tuberosum <400> 17 atggcttctt ccacactctc tcatgtaact ccttctcagc tatgctcaag caaaagtggt 60 gtttcatcgg tttcacaagc gttgctcgtg aaaccgatga agattaatgg tcatggaatg 120 ggaaaggata agaggatgaa agtgaaatgt atggcagcta gcattccagc agatgataga 180 gtgcctgata tggaaaaaag gaatctcatg aatttgcttt tattgggtgc acctgcacta 240 ccaactggtg gaatgcttgt accttatgct actttctttg cacctcctgg atcaggaggt 300 ggtagtagcg gtaccattgc caaagatgct aatggcaatg atgttgttgt aactgaatgg 360 ctcaaaactc atgcacctgg aactcgaacc ctcacacaag gactaaaggg agatcctact 420 taccttgttg tggagaacga tggaacactc gcgacctatg gtatcaacgc cgtgtgcact 480 ccccttggtt gtgtcgtgcc atggaatact gctgagaaca agttcatttg cccttgccat 540 ggatttcaat acaacaatca aggaaaagtt gttagaggac ctgctccttt gtccttggca 600 ttggctcatg cagacattga tgatgggaag gtggtgtttg ttccatgggt tgaaacagac 660 ttcagagctg gggattctcc atggggggct tag 693 <210> 18 <211> 885 <212> DNA <213> Solanum tuberosum <400> 18 atgcaaccaa ctaaggttgg tggtgtctct gccagaaaca acttgcaatt gaggtcatct 60 caaagtgtga gcaaggcatt tggtcttgaa ccatctgcat ccaggctttc ttgctctttg 120 caaactgatc ttaaagattt tgctcagaaa tgtactgatg ctgccaagat tgctggattt 180 gcccttgcca cctctgccct tgttgtctca ggagctaatg ctgaaggagt tccaaaacgt 240 ctaacctttg acgagattca gagcaagaca tacatggaag taaagggaac tggaactgcc 300 aaccagtgcc ctaccataga tggaggtgtt gatagctttg ccttcaagcc aggcaaatac 360 aacgccaaga agttctgctt agagccaaca tcattcaccg tcaaggcaga gggcgtaagc 420 aaaaattcag ccccagactt ccaaaagacc aagcttatga ctcgcttaac ctacactttg 480 gatgagattg agggaccctt tgaagtatct cctgacggca ctgttaagtt tgaggagaag 540 gatggaattg attacgctgc tgttactgtc cagcttcctg gtggtgagcg tgttcccttc 600 ctcttcacca tcaaacaact tgtggcatct ggaaagcctg agagcttcag tggtgacttc 660 cttgtgccat catacagagg ttcatcattt cttgacccaa agggcagggg gtggatccac 720 tggttatgac aatgctgttg cattgccagc tggaggcaga ggagatgagg aggagctcca 780 gaaagagaac gtaaagaata ccgcgtctct acaggaaaga tcacatttac tgttaccaag 840 agcaacccac agactggtga ggtgattgga ggtggtttga agagc 885 <210> 19 <211> 432 <212> DNA <213> Solanum tuberosum <400> 19 atggagagag ttaacaagtt gtgtgtagca ttttttgtca tcagcatgat gatggcaatg 60 gccgcggcgc agagcgctac caacgttagg gcaacctatc atttgtacaa tccgcagaac 120 ataaactggg atttgagaac tgctagcgct tactgcgcta cttgggatgc tgacaagcct 180 ctagcgtggc gccagaggta tggctggacc gctttttgtg gtccagctgg acctaggggc 240 caagcttcat gcggtagatg cttgagggtg acaaacacag gaacaggaac acaagaaaca 300 gtgagaatag tagatcaatg cagcaatgga gggcttgatt tggatgtaaa cgtgtttaac 360 cgattggaca ctaatggatt gggctaccag aggggcaacc ttaatgttaa ctatgaattt 420 gtcaactgct aa 432 <210> 20 <211> 360 <212> DNA <213> Solanum tuberosum <400> 20 atggctactt cttcaatagt ttcaatggca attccactaa cttgcaaaag ccaaaagaaa 60 caaccaattg ttgaaggttt tttcaagcca ttaccaatta ggccttcaaa acaaattgca 120 acttcaaaaa agtttgtagt catcaaggcc tcttcaatta aggaaaaagc agtggtaggg 180 ttgacaacag gtgtactcac agcatcaatg gttgtacctg atgtagcaca agctgctgag 240 agtttgtcac catcacttaa gaacttctta cttagcattg tgtctggtgg agttgtgctt 300 ggtgctatta ttggtgctat tattggtgtc tccaactttg atcctgttaa gaggggttaa 360 360 <210> 21 <211> 399 <212> DNA <213> Solanum tuberosum <400> 21 atggcgttaa gggaaacgat gctacgaacc gccgggaagt tcgccggtgt gaaccctaat 60 ccccgacgat ttatccacat cacttcacgt ccggcgcctc tagatagcgt ggttgatcgg 120 ctgacggcga catcaccgtc gttggtttta cctgaaaatg agaataacag caatatgttt 180 ccaaattcac ctttcaattg ggggtccatg gagcttatgg ctgttcctaa gaagaaggtt 240 tctcctcaca agagaggcat aaggaatgga ccaaaggctc taaaacccgt tccagtgatt 300 attcgttgca aggcttgtgg tcgagtcaag ttgccacatt tcttttgttg cagtgggatt 360 aagcagaacc ctggtgaaga gaacaattct accggttga 399 <210> 22 <211> 513 <212> DNA <213> Solanum tuberosum <400> 22 atggccactg ttacctctgc tgctgttgct attccatctt tcactggcct caaggctggt 60 gcttcctcat cttccagagt tagcaccgct gcatccgcca aggtggcagc tgccccagct 120 gccagattga ccgtgaaggc ttctttgaaa gatgtcggtg ctgttgttgc tgccaccgct 180 gttagcgcga tgcttgctag taatgccatg gcacttgatg tgttgcttgg tggtgatgat 240 gggagtcttg cttttattcc tggaaacttc agcgttagcg ctggtgagaa aattacattc 300 aagaacaatg cagggtcccc acacaacgtc gtattcgatg aggatgaaat cccagctggt 360 gtagatgcaa gtaagatttc catggctgaa gaggatcttc tgaatgcagc aggagagaca 420 tatagtgtca ctttgagtga gaaaggaact tacactttct actgtgcacc tcaccaggga 480 gctggaatgg ttggcaaagt tactgttaac tga 513 <210> 23 <211> 735 <212> DNA <213> Solanum tuberosum <400> 23 atgcgtttaa gctgtaatgg ttgccgggtt ctccggaaag gttgcaatga aaaatgctcc 60 attaaacctt gtcttcaatg gatacaatct ccagaagctc aatccaacgc cactcttttc 120 ctcgctaagt tttatggccg tgctgggctt ctcaatctca tcaattctgg acctgaccat 180 ttgcgtcctg ctattttcag atcgttgttg tatgaagctt gtggccgtat cgtaaatcca 240 atttacggtt ctactggttt gttaacttcc ggcaactggc ctctctgtca agctgctgtt 300 gaagccgttc taaacggtgc gccgattgtt aaactttcag tcgactcggc tgcagatgca 360 acgagccctt cgcttaaagc atgtgatatt cgccacgtgt ctagagaaaa taactcggat 420 cccgatctgc acaaagttaa gtcgagaacc cggtttaagc gggtatcggc taaaccggag 480 ccgaggaagc agtcgccgga tgttgacgat gcggctaggg tgatgtggag ttggtgtcat 540 aaggaggagg aagaggctgt cgggtcgccg agccttaatt cgggtctgag ccaacagcgg 600 acggaagagc cgagccggga aggagaaagc ggtgatgtgg atagcggctc ggtggagact 660 gttgaagcct ttccagcttc tagcgatgtt ggcttggaac taacgctggg gttgctaccg 720 ttgatgagaa gctga 735 <210> 24 <211> 177 <212> DNA <213> Solanum tuberosum <400> 24 atgtattttt ttagatttct tctttcaatg gtttttggtt gtcttgtcta ctctagttat 60 ttctttttct tcggattgga tctacataat tttttcgata ttttcttcgg gttgctaact 120 caacggcaga gtactcggct tttaagtgcg gctagtgtct tttacacata tggatga 177 <210> 25 <211> 642 <212> DNA <213> Solanum tuberosum <400> 25 atggctactg cagcatctac aatggctagc cagttgaaga gcagctttgc ttcgtcttta 60 accagaggtc acggccttgt tactcccaaa ggcatttctg gtgcaccttt taagattttc 120 ccttcaacaa gaaagtcttg cttcaccgtc aaggctgtcc aagctgataa accaactttc 180 caagtgattc agccccttaa tggtgaccct ttcattggaa gtcctgaaac tccagtcacc 240 tcaagtccat tgattgcatg gtacctctcc aacttgcctg cctaccggac cgcggtggac 300 ccacttctcc ggggagtaga agtgggcctg gcccatggat tcttactagt tggcccattt 360 gtaaaggctg gtccgttaag aaacacagac tatgcagggg gagctggttc tttggcagca 420 gcaggacttg ttgtgattct tagtatgtgt ttgacaattt atggaatttc atcattcaaa 480 gaaggagatc catcaacagc accagcatta acattgactg gcaggaaaaa agtgccagat 540 caattgcaaa ctgcagatgg atgggctaaa ttcactggtg gattcttttt ttggtggaat 600 ttctggtgtc acttgcgctt atttcctcct atatgttctt ga 642 <210> 26 <211> 318 <212> DNA <213> Solanum tuberosum <400> 26 atgggtgccg gtttcctcgt tggcgtcatc ggcgttctaa ttctatctca tgctacctat 60 tccacaatcc aatatagggc tttgcttaag attactgagg aggagttttc tggaccaccc 120 attgatgttg ttatcgaatt gatagttagt ttagtgttat gcttgtgggc tgcaatggat 180 gcccctggaa atttcaagtc aatccatcct caatctgagg aaaacagggt ggtcgctttg 240 cctgccaatc tggattttat gatctttaat caccgtggga aaatatttcc tctggaaacc 300 aagttgaagt tgaagtga 318 <210> 27 <211> 382 <212> DNA <213> Solanum tuberosum <400> 27 atgcttaatt gtgatggcgc tgaacttgaa gcaggcatca atattcccaa tccatgttgc 60 tagtgtaact gggtccacga ggaaaggagt gcaaaaggtg ttgatgaatg gatcaggatc 120 agcaggagta ccaaagtata aggggacgca gatgagagag aaacagctga cggaaatgat 180 agagaagaaa gtgaaggaag ctaaggaggt gtgcggtgaa gacgccaggt cggacgagtg 240 caaggtggca tgggatgagg tggaagaagt aagccaggct aaggcccatc tgcgtatcaa 300 gcttcagctc aatcaagatc cccttgaacc ttactgtcaa gataaccctg acgccgacga 360 gtgccgcacc tatgaccatt aa 382 <210> 28 <211> 291 <212> DNA <213> Solanum tuberosum <400> 28 atgagttacc ttaaccgcgt ttgggtagcc acaaccgtcg ccgtcgtaaa cagccacgcc 60 gatcacggcc agaaattgaa atccggcatc caatctctaa accacggcaa aaaacgattc 120 tcttcttctg tcgcttgcgg cgccgacgtt gccggtctcc ggccgatgtc cggtatgtta 180 ggatctgacg tcggagggtt catcggaggt ggaaaaggcg aggagaagag gaaacaaact 240 gatgagtcac tccggcaagt tatgtaccta agctgctggg gaccaagcta a 291 <210> 29 <211> 681 <212> DNA <213> Solanum tuberosum <400> 29 atggcacaag tgaagttgct aggtttttgg tatagtccat ttactcacag ggttgagtgg 60 gctctaaaga ttaagggggt aaaatatgaa tatattgaag aagatcgata taacaagagc 120 tctttacttc ttgaattgaa ccctatttac aagaaagtcc cagtgctcat tcacaatggc 180 aagcccatta ttgattctat ggtcattctt gagtacatcg acgagacatt tgaaggccct 240 tccatcttgc ctaaagactc tcatgaccgt gctttagctc gtttttgggc taagttcctt 300 gatgataagg tggttacggt ggtaaatgca ttcttaggca aaggagagga gcaagagaaa 360 gctaaagagg aagtttgtga gatgttgaaa attcttgata atgagctcaa agacaaaact 420 ttctttgtgg ctaacaaatt tggatatgct gatattgttg caaatttggt tggactttgg 480 cttggagttt tccaagaagg ctttggaatt gaattagtaa caagtgaaaa atttccaaat 540 ttttgtggtt ggagagatga gtacgttaag tgcagtcaag tcaaggaata tctaccttcg 600 agaaatgatt tgcttgcttt ttttcaagtt ttcgttcgtg ctcaagttgc agctgcagct 660 tctgcttcta ctaaaaaata a 681 <210> 30 <211> 702 <212> DNA <213> Solanum tuberosum <400> 30 atgaaagtac actcctgcat tcttcctctt ccctgcactc cttccaccgg aattttggtc 60 tccggcggtc gtacggaagc taaccctgct gctaacaccg gagagctata ttcattaact 120 ttgaattcct ctaattatac cccatttcat tataaagatc tgaaattttc cccagacttc 180 tctatggatt tttcgataat ggcaattggt tcgaatgtga gtgaagatac ggatgacatg 240 tttgatgatc tcctaagaac gcaagggaaa gtggtattca aaagaaatga ccaaaaacct 300 gcaagtgagg agattgatga tgatgctgaa agcctatcct ttgcaagggc tgtggctaaa 360 gttgccagtg atgtcgaggc tgaagatata aaggttctct ttgtcaagcc tctactatat 420 tgcactcggt ttttcattat tgctactgca tgctctcgcc cacatattga tgccattggg 480 accagagtaa gagatatggc tgaggatcaa tatggaagag ttgcttctgg agattcaaaa 540 ctcaactcgt ggacactact agactttggt gacgttgttg ctcgtctatc tcctcctccg 600 cagcgtgagt acgaccacct ggaataattc tctggctatg cagcatccat cgtgctacca 660 tttgagaacc accaacaatt gcgaggacct actggatatt ga 702 <210> 31 <211> 447 <212> DNA <213> Solanum tuberosum <400> 31 atggcttcca agcggatctt gaaggagctc aaagatcttc aaaaggatcc tcctacttct 60 tgtagtgctg gacctgttgc tgaggacatg tttcattggc aagcaacgat catgggtcct 120 ccagacagtc cttattctgg tggagttttc ctggtgacaa tccattttcc tccagattat 180 ccattcaagc cacctaaggt ggctttcagg acaaaggttt tccacccaaa cataaacagc 240 aacggaagca tatgccttga cattttaaag gagcagtgga gccctgccct aacaatttcc 300 aaggtgttgc tttcgatatg ttcacttctg acggacccta atcctgatga tcctttggtc 360 ccggagattg cacacatgta caagacagac cggaacaagt acgagagtac tgcaaggagc 420 tggacccaga agtatgccat gggctaa 447 <210> 32 <211> 1182 <212> DNA <213> Solanum tuberosum <400> 32 ggctgctctt ctcttttaca aaaaccctaa taccttctcg ccgtcagcaa tggctaccgc 60 tgccgccatc ccaacgacca ccgtccaatc gttcgaagat gacatggcta ctgacggtgg 120 ctccgccgca aactccgttc ctcttcccgc tgtgatgaag gctccgatcc gtcccgacgt 180 tgtcacttac gtccacgcca acatttcgaa aaacgcccga caaccttacg cagtgtcgaa 240 aaaggccggt caccagacct cggctgagtc atggggtacc ggtcgtgccg tttctcgtat 300 ccctcgtgtt cctggtggtg gtactcatcg tgctggacaa gctgcttatg gtaacatgtg 360 tcgtggtggt aggatgtttg ctccgactaa aatctggcgc cgttggcacc gtaagatccc 420 ggttaaccag aagcgtcatg ccgttgtttc tgccattgct gcatctgcag ttccttcact 480 ggtcctagct cgtggacatc gtattgaatc agttcccgaa cttcctctcg ttgtttctga 540 ttcagttgag agtattgaga agacttctaa tgcaatcaag gctctgaagc aaattggtgc 600 ttatgctgat gctgagaaag ctaaggatag tcatgctatt cgccccggaa aaggtaaaat 660 gcgtaaccgt aggtacattt ctcgtaaagg accactcatt gtttatggaa ctgaaggggc 720 taagcttgtt aaagcgttcc gtaatctccc tggagttgaa atctgtcatg tggatcgatt 780 gaatctgttg aagcttgctc ctggaggtca tttgggaagg tttattgtat ggactaagtc 840 tgcatatgag aaattggatg agatttacgg tacatttgat aagccatctc agagaagagg 900 ggatatctat tgcctagggc cagatggcca atgctgattc ttgctaggat catcaactct 960 gatgagttca atctgttgtg cggccatcag aggacgcgag agcacattga gaagaatcct 1020 ttgagacctg acggctgttg gagcttatcc atatgcaaac acacaagaga ggagttctct 1080 ttgtctgagc caagagtcag gctgaaggaa acttgcaaga aagctcaatt ctaggactcc 1140 tagtcagcag tgtcgacaga tcgattcgca tcactgaggt cg 1182 <210> 33 <211> 527 <212> DNA <213> Solanum tuberosum <400> 33 gtggtggtga ttgcacccca tgcagaggca ttaacttgtg gtcaagttac agctggcctg 60 gctccttgcc tcccttatct tcaatggccg cggtcctcta ggaggctgtt gcagtggtgt 120 taagggtctg ttgggttcag ccaagactac cgcggatcga aagacagcat gcacttgctt 180 gaaatcagcc gctaatgcta taaagggaat tgatttgaac aaagctgccg gtatccctag 240 tgtttgtgga gtcaacattc cttacaagat cagcccctcc actgactgtt ccacggttca 300 gtaaggtaga tcatcagaat atggaaccat gaggagaatt aagaataaga tgaatggagt 360 gtgatcctgc ttatggatcc attatttatg tgttgtgtct gttttctctt tcttttttcg 420 ttgtgttttt gggggttttg tatcgaaatt tcacgattgg ttgtattacg ttaatccaat 480 cttgataata tatctatctt atttagtaaa aaaaaaaaaa aaaaaaa 527 <210> 34 <211> 1217 <212> DNA <213> Solanum tuberosum <400> 34 ggtagcaaaa atggaaattg aaatcttatg cacaaagctc ataaagccat cttcacctac 60 tctctctcac aatcaatgtt acaagctctc tttctttgat caaatagctg agagagaaca 120 ccttcctgtt gttcttttct atccttataa taatttcaat agccccacta tcgatgaaca 180 gcttgagaaa tccctttcta atgtattaac tcatgtttat ccagcagcag gaaggtatga 240 caaagatgaa tgttcgattc tttgccttga tcaaggtgtt ccctacacta aggccaaggt 300 caattgtaag ctggataatt tcttggagaa agcacacaag gatcttagtc ttgcagcttt 360 attttggcca catgaaaaca agaatgtgga tcaaaataat tttatggtat caccaattgt 420 caccattcaa gtaacaaaat ttgaatgtgg tggcctagct ttatcattca gtgtttcaca 480 ccctgcaatt gatggtgtca catgttttaa ttttctgttt ggatggggaa aagtgtgtag 540 attgggaact ccaattgata agattaattt tctaagcttc aatttaggtc atgtttttcc 600 aaccagagat atatcaagtc ttttcaagtc aatccatgta gaaaatagag aagaaaatat 660 tgttgttaag agatttgtcg tccatgaagc tgctctatca aggctcagaa aacaatgcat 720 tgatgaatca ggtggagctt tgaattttca accttcaagg gttgaaatta ttacagcaat 780 cctatgggag ggctttaatc cgcatttcag cagctagaaa tggatatttc aggtcttctc 840 taatggactt accattgaat ttgcgctcta aatcatcttt acctcaagta aacaactcta 900 tgggaaattt tagaatcgat attccgataa aaattcattc caggggagac caggatggaa 960 ttacacaact tcatatattt atcaggatac tggtgaataa gttgttgctt cttgtaccaa 1020 cgccgtccca gatgaatatt gtcgacatta gtttatataa tatacgaagc cttgagtccg 1080 gcaatgggag ggtatacaca gaattccaat gggcaggtcc atgtctcctg aatggtgcca 1140 ttggaactcg aggaagtctg gcatatttag gtgggacagc gaattcaggc ctctacgtaa 1200 aaaaaaaaaa aaaaaaa 1217 <210> 35 <211> 969 <212> DNA <213> Solanum tuberosum <400> 35 gatgctcgcc gggattctga aaacgaacgg cgtacaagcc gacggaatca attttgacaa 60 gggtgccagg acggctttgg cgttcgtgac tctacgcgcc gacggagagc gtgagtttat 120 gttttacaga aatcccagtg ctgatatgtt gctcacgccc gatgagttga gtcttgatct 180 tattagatct gctaaggtgt tccactatgg atcaattagt ttgatcgtgg agccatgtag 240 atcagcacat ttgaaggcaa tggaagtagc aaaggaggca ggggcattgc tctcttatga 300 cccaaatctt cgtttgccgc tgtggtcttc ggaagaagag gccaggaagc aaatcaagag 360 catctggaac tatgctgatg tgatcaaggt cagcgatgtg gaactcgaat tcctcactgg 420 aagcaacaag attgatgatg aatgcgccat gtccttgtgg catcctaact tgaagctcct 480 cttggtcact cttggtgaaa agggttgcaa ttactacacc aagaaattcc atggaagcgt 540 tggaggattc cacgtgaaga ctgttgacac cactggagct ggtgattctt ttgttggtgc 600 ccttctaacc aagattgttg atgaccaagc cattcttgag gatgaagcaa ggttgaagga 660 agtactaagg ttttcatgtg catgtggagc catcacaacc accaagaaag gagcaatccc 720 agctctgcct actgtatctg aagtcctcac tttgctcaag ggaggagcat agaaacatca 780 tgttatcttt ttattttttt tttccatctc catatattcc cccctctatg agttttttta 840 ctttcaagct agttcaattt ttagtttggg attttaatgt tttgttgtga tgacggtcca 900 tcaagcactt aataaactaa gctttcttcc aaaaaaaaaa aaaaaaaatg gtgttctcat 960 cgtaccccg 969 <210> 36 <211> 553 <212> DNA <213> Solanum tuberosum <400> 36 cataaggatt taaaattccc aggaagcata cagtctagtc tccatgcgaa gcagttgatg 60 tacagattgt tgcacagaga tcccaaaaac aggttaggat ctcgtgaagg agcaaacgaa 120 atcaagcagc atccgttctt ccggggtgtg aattgggcat taattcgttg catgaaccct 180 cccaaactag attctgctcc ttttttggga gctgaagctg agaaagaagg aaaagacatc 240 aatcctgaga tggaggattt acaaacaaat gttttctaaa agaaatgaaa atatttatat 300 cccaatcaaa tcaagtgcat gagtggagaa aagtggtctt cagaccctcc tagcaacagt 360 taattacttg ttgatttgta taaaataaaa tgatggttgt tcatttgttt tttttcttcg 420 tttgatggtt tgtgttgttc cttttatatg catagtttgg ttgttttgta attcttaatt 480 catgaattcc tcagtagatg ttgagttaat taatttgtta agcctaattt tctaaaaaaa 540 aaaaaaaaaa aaa 553 <210> 37 <211> 862 <212> DNA <213> Solanum tuberosum <400> 37 gaattttaaa aaactagcaa ctctatctct tccaatttat agagagagag agagagagag 60 agagagagag aaaatgacgg ggttggtaat ggtgacaagg ggaggtgggt gtggaaatgg 120 gaagaacagt tcaaaggcgg cagctgctga aatgcaaaat cagcagcagt tgtctttggt 180 ggatttcata ttagcagctc taaggaaatc aatggtgtct tgtcgtgtag atcgacaaga 240 agatgttgtt tctgctgtgg aaatagggtg gcccaccgat gtccagcatc tcacccatgt 300 cactttcgat cgatttcatg gctttctggg tcttcctctt gagtttcaag tcgaaatccc 360 atgtagagta ccaagtgcca gcgtcagcgt gtttggtgta tctgcagagt caatgcagtg 420 ttcttatgat acaaggggca acagtgtccc aactattctc ttgttaatgc aagaacgtct 480 atactcacaa aatggtctta aggctgaagg aatcttccga attaacccag agaatagcca 540 ggaggagcat gtaagggacc agctaaatag aggcatcgtg cctgaagaca ttgatgtttc 600 attgtttggc tggtctcatc aaagcctggt tctgagaact gccatcaggt gtgctttgat 660 gggctttcac cagaacaggt tttgcaatgc aacaccgagg aagaatttgt tgagcttgtg 720 aacagcttaa aaccaccgga gactgcattg cttaacgggg caattgatct tttggctgat 780 gttgttgaac agaggaatcc acaaaatgaa tgccagaaat attgaatggt ttttgctcca 840 acttgaccca gatgtctgat cc 862 <210> 38 <211> 917 <212> DNA <213> Solanum tuberosum <400> 38 cataaaacaa ctgaatgata tataccgcta caaaacagaa gagtactctc acaccgcagt 60 aaacaagttt aatgtgatgc cagattccct tccggagtgg gtttttgatt tcatgccaac 120 tcgtggtggt tacttcatcg gaaatgttag tcctgctcac atggacttcc gttggttttg 180 tttgggaaac tgtatttcaa tcttgtcatc tttggctaca cctgagcaag cttccgccat 240 aatggatcta gttgaatcaa gatggcaaga gctagttgga gaaatgccgc tgaaaatctg 300 ttatcccgct atggaaggtc atgaatggag aattgtaact ggatgcgatc ctaaaaacac 360 tagttggagt taccacaatg gtggaacttg gccagttctt ctgtggctcc ttactgcagc 420 agctatcaag actggtcgac cccaaatagc acgacgggcc attgaactag ctgaatcgcg 480 tttactaaaa gatagctggc cagagtatta cgatgggaag cttggtcgat ttatcggaaa 540 gcaggcccgt aagttccaga catggtccat tgctggttac ttggtagcta gaatgatgct 600 ggaagatcca tctcatttgg gtatgatatc acttgaagaa gataaacaga tgaagcctac 660 catgaaaaga tctgcttctt ggacttgtta atatccttgt acacaatttt tctttcgaat 720 gccatgtttt tttttctttt tcttttttgc aattcgtaaa cattctcttt tagtttgtcg 780 caaaaatgtt gaattttcta tgaattggct acgagattgt atacttacct cgttttttcc 840 aattaatctg gttttcgtct agaaaaaaga aataaaagag agaagagtaa aaaaaaaaaa 900 accaaaaaaa aaaaaaa 917 <210> 39 <211> 436 <212> DNA <213> Solanum tuberosum <400> 39 gacatccaat tcaaatataa tttcaatatt caaactttca ctttacctta aagttgttga 60 taatttcttt gattgaaaaa aagaagaaga aatatggctg atcagtacga acagaacaag 120 ccatcagttg aagagactgt tggtgccaac gtggaggcta gtgatcgtgg tgtgtttgat 180 ttcattgtga gtaaaagaga ggaaaaacca agtcatgctc atgaagaaga ggcaatttca 240 tctgagtttt gtgagaaagt taaagtaagt gaagaagaag aacacaagga ggaaaagaaa 300 gaagagaaga aacttcatcg atcaagtagc agctctagta gctcgagtga cgaggaggaa 360 gaaattggag aggatagaca gataaccaag aagaagaaaa agaagggatt gagggaaaaa 420 aaaaaaaaaa aaaaaa 436 <210> 40 <211> 764 <212> DNA <213> Solanum tuberosum <400> 40 tgattaccct gtcccacgca aatcgtttac ctgtaactat tcaatacccc tacgaaaaat 60 tgatcacatc ggaacgtttc cgaggccgaa tacactttga atttgataaa tgcattgctt 120 gtgaagtatg tgtgcgtgta tgtcctataa tgttacccgt tgttgattgg aagttggaaa 180 ctgatattcg aaagaaacga ttgcttaatt acagtattga ttttggaatc tgtatatttt 240 gtggtaattg cgttgagtat tgcccaacaa attgtttatc aatgaccgaa gaatatgaac 300 tttctactta tgatcgtcac gaattgaatt ataatcaaat cgctttgggt cgcttaccaa 360 tgtcagtaat tgatgattac acaattcgaa caatttcgaa ttcacctcaa ataaacaatg 420 aataaaccct taattcgatt caaaaaaatt acgaattacg aaggcttagt tcggatctaa 480 aacaatgaga tttttgcttg ggcaattcaa actagtggtt gcttaatgag actcctagct 540 taatttagat tgaaaacaaa accgatttca atattatata ttataatagt aatggtttca 600 aagtagataa atgatatcaa aaatagatag gtttaaacta ctctttcgat gaataaagaa 660 tggatagtgg tccaataaag taaaagcatc tacgtcaatt catacccatt agactttcat 720 tcaattaaca atttcaaagt atcataaaaa aaaaaaaaaa aaaa 764 <210> 41 <211> 687 <212> DNA <213> Solanum tuberosum <400> 41 atggctctta agtcctttgt attaatcatt gccatattgg ctgtggttac ttcaataagc 60 catgcatctg atcctagtcc tttacaagat ttttgtgttg ctgttaatga ctccatgact 120 actgttttcg tgaatggaaa agtatgcaaa gatccaaggg ttgtcactgc tgatgacttc 180 ttcaaatcag gcttaaacgt agctggaaat acttcaaata atgttggatc tgcagtaact 240 gctgtgaatg ttaacaactt accagggctc aacgctctgg gcatttcatt agttcgtatt 300 gactatgcac catacggtct caacccacca catacacacc ctagaggaac cgaggttctt 360 acggtccttg agggcacact ctatgttggt tttgtccttt caaacccggg tccaaacatg 420 aagaacaagc tctttaccaa gattctacat cctggagatg tgtttgtttt cccaataggt 480 ctcattcatt ttcaattcaa cgtgggaaaa actaaggctg ttgcatttgc tggactcagt 540 agtcaaaatc caggagtcat cactattgca aatgcagtat ttggctcaga ccctccaatc 600 aatgacgata tcctcgctaa agcatttcaa gttgataaga aagttgtgga ttatctccaa 660 tcacaattct ggtgggataa caactaa 687 <210> 42 <211> 345 <212> DNA <213> Solanum tuberosum <400> 42 atggaaatgt ttggcaaaat tgcatgcttt gtgcttttgt gcatggtggt ggttgcaccc 60 cgtgcagagg cactgagctg cggcgaggtt acatctggct tggctccctg cctcccttat 120 cttcaagggc gcggtcctat tggagggtgt tgtggtggtg ttaagggtct gttgggtgca 180 gccaagaccc cagaagaccg gaagacagca tgcacttgcc tgaaatcggc agctaattct 240 attaagggca ttgatacggg aaaagccgct ggtctccctg gagtttgtgg agtcagcatt 300 ccttacaaga tcagcccttc cactgattgc tccaaggtcc agtaa 345 <210> 43 <211> 831 <212> DNA <213> Solanum tuberosum <400> 43 atggcagctg ctacaatggc tctttcttcc ccttcttttg ctggacaagc agtgaaactc 60 tcaccatctg cctcagaaat ctctggaaat ggaaggatca ctatgagaaa ggctgtcgcc 120 aagtctgccc catctagcag cccatggtat ggccctgacc gtgttaagta cttgggcccg 180 ttctctggtg agtccccaag ctacttgact ggtgaattcc ctggtgacta cggctgggat 240 accgctggac tttcagcaga ccctgaaact tttgccaaga atcgtgaact tgaggtgatc 300 cactgtagat gggctatgct tggtgctctt ggatgtgact tccctgagct cttggcccgt 360 aacggtgtca agtttggtga ggctgtgtgg ttcaaggccg gatctcagat cttcagtgaa 420 ggtggacttg attacttggg taacccaagc ttggtccatg cacagagtat cttggccatc 480 tgggcttgcc aagttgtgtt gatgggagcc gttgaaggat accgtattgc cggtggacct 540 cttggtgagg tcgtcgaccc actctaccct ggtggaagct ttgacccatt aggtcttgct 600 gatgatccag aggcatttgc tgagctcaaa gtaaaggaga ttaagaacgg tagacttgct 660 atgttctcta cgtttggatt cttcgttcaa gccattgtta ccggaaaggg tccattggag 720 aaccttgctg atcaccttgc agacccagtc aacaacaacg cctgtcctac gccacaaact 780 ttgtccccgg aaaatagaat cttaaaacat tctcagtttc tggtcgtttg a 831 <210> 44 <211> 465 <212> DNA <213> Solamun tuberosum <400> 44 atgtcactga tcccaagaat cttcggcgat cgaagaagca gcagcatgtt tgatccattc 60 tcaatggacg tgttcgattc gttcaaggaa ctaggcttcc cagtttccaa ttcaggagag 120 acctctgcat ttgcgaacac tcgagtcgac tggaaggaaa ctccagaagc tcatgtgttc 180 aaggccgatc ttccagggct taaaaaggag gaagtgaaag tggagatcga agaagatagg 240 gttcttcaaa ttagcggaga gaggaatgtg gagaaagaag ataagaatga taagtggcat 300 cgcgtggaac gcagcagtgg gaaattcatg agaagattta gacttccgga gaatgcgaag 360 atggatcaag ttaaggcttc gatggagaat ggagtgctta ctgttactgt tccaaaggaa 420 caagtgaaga agcctgatgt caagtccatt gagatctctg gttaa 465 <210> 45 <211> 594 <212> DNA <213> Solanum tuberosum <400> 45 atggtgcacg tctcttttta tcgcaactat gggaagacct ttaagaagcc acgaaggccc 60 tacgagaagg agcgattgga tgcagaattg aaacttgtag gagagtatgg actaaggtgc 120 aagagggagc tttggagagt tcagtatgct ttgagccgta tcaggaatgc tgctagaatg 180 cttctgacat tggatgagaa ggattcacgt cgtatttttg aaggtgaagc tctcctgagg 240 aggatgaaca ggtatggatt gttggatgag agccaaaaca agcttgatta tgtcttggca 300 ctcaccgtgg agaactttct tgagcgtcgt ctccaaacac ttgtcttcaa gactggcatg 360 gctaagtcaa ttcatcatgc cagagtgctc atcaggcaaa ggcatatcag agttggtagg 420 caggtggtga atgttgcttc ttttatggtg agagtggact cccagaagca cattgacttc 480 tctctcacta gtccgtttgg tggtggccgt cctggaagag tgaagcgaaa gaaccaaaag 540 tccgctgcca agaaggctgc tggtggtgat ggtgatgaag aggatgaaga atga 594 <210> 46 <211> 915 <212> DNA <213> Solanum tuberosum <400> 46 atgactatac tcattgagca gcttcctgtt gagatccagg cagatgaaac gaatattcat 60 catgctcaga atcaaaagga attggtattg gttgatggat ttgcagttcc tcaaaccaat 120 gcatttggcc ataattttag ggactacact gttgaaagtg caaggcaaca aggggtagaa 180 tttttttaca agatcaatca ctttaatcaa acctatgatt atgtaagtaa gatgagggtt 240 gaatatgcaa aattggacaa ggcagaaatg agcatctggg aatgctgtga actattgaat 300 gatgtggttg atgacagtga ccctgatttg gatgaacctc aaattcaaca cttgttgcaa 360 agtgctgaag ccattaggaa agactatcct aatgaagatt ggctccattt gactgccctt 420 attcatgatt tggggaaagt gcttcttctt cctagctttg gagagcttcc tcaatgggct 480 gttgttggtg acacgttccc tgttggctgt gctttccatg actcaattgt tcactccaag 540 tactttaagg aaaatcctga ctacaataat cctgcttaca acagcaaatt tggagtttac 600 tctgagggtt gtggactaga caaagtgttg atgtcatggg gtcacgataa ttatatgtac 660 ttggttgcaa aagagaatgg gacaacgttg ccatcagctg gtcttttcat cgttagatat 720 cattcatttt atgccctgca taagtctgga gcttacaagg aactaatgaa tgaggaagat 780 aagaaaatct taagtggctt catattttta caatatgact tgtacagcaa agcaagttca 840 gttaatgtgg agagtcagcc ttactacatg tctctaattg aaagtatttc cagcaagctg 900 aaatgtgaga gctga 915 <210> 47 <211> 559 <212> DNA <213> Solanum tuberosum <400> 47 atggtgacag tggagtctga tttcgttgta atactagctg cgttgctctg tgcgctgatt 60 tgtgtagttg gtctcatcgc cgtcgcgaga tgtgcgtggc tccggcgagg aatcggcgcc 120 ggcgggatcg gtggtagtca accgtcggcg aataaaggat tgaagaagaa agtgttgcaa 180 tcgttgccta aattcacgta cgatccaagc agtacggcga atggtgcggc gttcacggcg 240 gagtgtgcga tttgtctggc ggagtacgcc gtcggagatg agatccgtgt gctgccgcag 300 tgtggacata ttttccatct tcagtgtatt gatacttggc tgggttcaca ctcttcatgt 360 ccttcttgtc gtcaaattct cgtcgttact cggtgccgga agtgcggtga gctcccggca 420 gtttcctgta aatccgacgg agctacggtt acaactccgg cggagtctcg ttcatgtggt 480 acatctatta gctcaaataa tttcttggtc taaagtaaca aaaaaaaaag gaaattagct 540 tttaagttat atacgctga 559 <210> 48 <211> 687 <212> DNA <213> Solanum tuberosum <400> 48 atgtctctta ctattccaac aaacctttca aaacccattt caacatctaa gtttgggtct 60 caatttacca caaaaccaag aacttcagcc attgtgtgtt cttcaaacca aacccctgtt 120 tctaattcaa cagaggaaag ttcatcttca ttaaaggcat tctctgcagc acttgctttg 180 tcttctattc tgttgtcagc accagttctt ccagcttctg ctgatatctc tgggcttaca 240 ccttgcaagg attcaaaaca gtttgctaaa agggagaagc aacagatcaa gaaacttgag 300 agttctttga aactttattc acctgatagc gcccctgcac ttgctattaa tgccactatt 360 gaaaaaacta aacgcaggtt tgacaactac gggaaacaag gactgttgtg tggatcagat 420 ggactaccac acttgatagt gagtggagac cagaggcact ggggtgagtt catcacacca 480 gggatcctct tcttgtacat tgctggatgg attggatggg taggaaggag ctacttgatt 540 gcagtaagag atgataagaa accaacaatg aaggaaatca tcatcgatgt tccattggct 600 aatagactcg tttggagagg cttcatttgg cctgttgctg cttacagaga gtacttgaat 660 ggagacctca ttgaccctaa tgtctaa 687 <210> 49 <211> 438 <212> DNA <213> Solanum tuberosum <400> 49 atggcacaat acggcaacca agaccaaatg cgcaagactg atgaatatgg aaaccatgtc 60 caagaaactg gaactcatca aggtaccggt atgggaggca tgcaggctgg tactggtggt 120 atgatggggg gcatgggtac tggcggtatg ggtggcactg atggaggata tggaacacgc 180 accggtatgg gaatgggtat gggtactggt acccatcacc atgaggggca acagcagctt 240 cgtcgatccg acagctctag ctcgtcggag gatgatggag aaggtgggag gagaaagaag 300 gggatgaagg agaagataat ggagaagatg ccaggaggag gacatggcca gcaggaagga 360 caaacaacag gtgaaggagc tggagagaag aaaggaatga tggacaaaat caaggacaag 420 atccctggga tgcattga 438 <210> 50 <211> 591 <212> DNA <213> Solanum tuberosum <400> 50 atgggtttga atatggcaat aaatctggat gtgcatgagt tggtggtttt tggggatttc 60 aatttgctca ttcgacaagc tcgaggcgaa tgggagactc gagacattaa actcatgccg 120 tacaaacaat gtttagaaaa cctcatcaaa aggttcaagt ccattgaatt caaatacatt 180 cccaggtttc agaatgagtt agctgatgct ttggccaccc tagcctcgat gcttccatac 240 ctgggtaata cctatattga cccgttggaa atccaagtta gggatcaaca tggttattgc 300 aatataattg aagtagagcc agatggtgag ccctggtatc acgacattaa acggtttcta 360 aaagctaaag aatatccaat acatgctgat agagataaaa aaagaactat taggcgactc 420 gccaattggt tcttcttgag tggcgatatc ttttataaaa ggactccaga tttgaatcta 480 ttgcgatgtg tgaatattca agaagcagaa acaactatga atgcggtgca tttaggggtt 540 tgtggccctc acatgaatgg ttatgttcta gcaaaaagat tatccgggta g 591 <210> 51 <211> 450 <212> DNA <213> Solanum tuberosum <400> 51 atggcagatc agctcaccga tgatcagatc tctgagttca aggaggcttt cagcctattc 60 gacaaggacg gagatggttg cattacaact aaggagcttg ggactgtgat gaggtcgttg 120 ggacagaacc caactgaagc tgagctccag gacatgataa atgaagtgga tgctgatggt 180 aatggaacca tcgacttccc agagtttttg aacctcatgg cgaggaagat gaaggataca 240 gactcagagg aggagctgaa ggaggcattc agagtgtttg acaaggacca gaatgggttc 300 atctctgctg ctgagctccg tcatgtgatg actaaccttg gtgagaagct tactgatgaa 360 gaagttgatg aaatgatcag ggaggctgat gttgatggcg atggacaaat taactacgat 420 gagtttgtta aggtcatgat ggccaagtga 450 <210> 52 <211> 660 <212> DNA <213> Solanum tuberosum <400> 52 atggacgcag gtgtagttgc tgcccccgcc ccggccgccg ccgccgccgc cgctaccgat 60 gcaagcaaag agaacaagat tcacactgat gtcatgcttt tcaatcgctg gagctatgac 120 ggagttgaga tcaatgacat gtctgttgag gattacatca cagcaactgc taacaagcac 180 ccagtttaca tgccacacac agctggtaga taccaggcca agcgtttcag gaaggctcag 240 tgcccaattg ttgagaggct caccaattct ctcatgatgc acggaaggaa caatggaaag 300 aagctcatgg ctgtccgcat tattaagcat gcaatggaaa tcattcattt gttgactgac 360 caaaacccaa ttcaagtcat tgttgatgct gttatcaaca gtgggccaag ggaagatgca 420 acacgtattg gttctgctgg tgttgtcaga cgtcaagctg ttgatatttc tccacttcgt 480 cgtgttaacc aagcaattta tttgctgaca actggtgcac gtgagagtgc tttcaggaac 540 atcaagacca ttgctgagtg ccttgctgat gaactcatca atgctgccaa gggttcttca 600 aatagctatg ccattaagaa gaaggacgag attgaaaggg ttgccaaggc caatcgttaa 660 660 <210> 53 <211> 360 <212> DNA <213> Solanum tuberosum <400> 53 atggggaaga ccttcaaaga tgaattttca tatgaggaaa gactcgcaga atcacgggat 60 ataatcgcca aatatcctga tcgagtgccg gtggtggctg aaagatattc aaaaactgac 120 ctccctgaga tggaaaagaa gaaatacctg gtacctcgtg atatgtctgt tggacaattt 180 atccacattc tgagtggcag gctgcatctg gctcctggaa aagctctctt cgtgtttgtg 240 aataacacct tgcctcaaac aacaagcttg attgagaccg tgtatgaatc ttccaaggat 300 aaagatgggt tcctctacat gtgctacagc agtgagaaaa cctttggtgg tcactcttag 360 360 <210> 54 <211> 285 <212> DNA <213> Solanum tuberosum <400> 54 atgatgatgt caattttcag ttcctttgat gccctctctg ctgaactttt tgggcagaaa 60 ttgagttttt ctaagccatc tagtactgac acaaaacaac aaggggttgg aactcttgta 120 tcagatcaaa aaaccgccgc ttctccgccg gctacttctt caaccggtat tctgaagaag 180 gccggagagg catcgccgcc gccatcttcg cggcagcaga agaggcagag gttcgcgctg 240 gaattcgacg gcgtacactg ttttgaaact ataattccct attga 285 <210> 55 <211> 417 <212> DNA <213> Solanum tuberosum <400> 55 atggcaagca cagtaatgag ctcattgagc ctaaagccaa ctttcacttt ggagaaaaca 60 tcagtgaaag ggcttccatc tcttgctaga tcttcttctt ccttcaaagt tgtggctagt 120 ggtgtcaaga agcttaagac tgacaaaccc tatggaatta atggaagcat ggccttgaga 180 gatggcgttg atgcctcagg caggaagccc aagggaaagg gtgtgtacca atatgttgac 240 aaatatggag ctaatgttga tggatacagt cccatctaca acacagatga gtggtctcca 300 agtggtgatg tctatgttgg aggtactact ggcttagcca tatgggccgt gaccttggtt 360 ggtattcttg caggaggtgc tctccttgtc tacaacacaa gtgctttggc acagtag 417 <210> 56 <211> 483 <212> DNA <213> Solanum tuberosum <400> 56 atgtcggacg aagaacacca ttttgagtca aaggcagatg ctggtgcctc aaaaacttac 60 ccgcagcaag ctggaaccat ccgtaagaat ggttacatcg ttatcaaagg ccgtccctgc 120 aaggttgttg aggtctccac ttcaaaaact ggaaagcacg gacatgctaa atgtcacttt 180 gtggcaattg acattttcaa tggaaagaaa ctcgaagata tcgttccatc ctcccacaat 240 tgtgacgtgc cacatgtcaa ccgtaccgac tatcagctga ttgacatctc tgaagatggt 300 tttgtctccc ttcttactga aagtggaaac accaaggatg acctccggct tcccaccgat 360 gaaagtctgc tgaagcaggt taaagatggg ctccaggaag gaaaggatct tgtggtgtct 420 gttatgtctg cgatgggcga agagcagatt aatgccatta aggatattgg taccaagaat 480 tag 483 <210> 57 <211> 897 <212> DNA <213> Solanum tuberosum <400> 57 atggcgaaca acgcaccgga gatatatgct gccgatgatt ttctcgagaa aattctggcg 60 attccctctt atgctagcct accggtaact gatataagcg ccggtgcctc atcggagaat 120 tcagcctctg gtatctctca gctccagcag cagcggctgt tccctttggg gttaagcttg 180 gacaatggct ttgctggtgc caacgacatt ggaggttttc aagtgaagac tgaaagagaa 240 gctatgcaca tggggaattc atattcaggt ctagagcatt tgcaatctca tgctgtttgc 300 ctaagtgttc ctcaagttca ccaagtccag ccttttcaag gccacccaac atcaagcaca 360 acagtaacaa taccacacca acctgcaatt catcctaggg ttcgggcacg aagaggacag 420 gccactgatc cacatagtat tgctgagcgg ttaaggagag aaaggatatc agaacgaata 480 aaggctttgc aagaacttgt ccccaactgt aataagaccg acagggcagc attggttgat 540 gagattctgg actatgtgaa gttcttaagg cttcaagtta aggtactgag catgagtagg 600 ctgggaggaa ctagcgcggt agcacaactt gttactgata ttccattgca gtctgttgag 660 ggagacagct gtgaaagtcg atccaaccag cacgtctggg agaagtggtc cgattctgaa 720 acagagcaag aggtagcaaa gctcatgggg gaagatgttg gaacagccat gcagtacctt 780 caatccaaat cactctgcat catgcctgtt tcacttgctg cacttatcta tcctactcag 840 caaactgacg accaatcaat ggtcaagcca gactacggcc ccatcgtaga cttctag 897 <210> 58 <211> 912 <212> DNA <213> Solanum tuberosum <400> 58 atggcagcaa accaaccgga agcctacgcc gatgatttcc tcgagcaaat tctcgctatt 60 ccttcctatg gcttgccggt tgctgatgtt ggcactccat cagagacgac gtcgtttacc 120 tcagcgtctg ccgtatctcg tcttaactct gccgctgctg ctggcctaca gcaacctatg 180 ttcccgttgg gattgagctt ggataacggc cgtgataacg tcagcgaagc tggtgcttat 240 gcagtgaagc atgaaagaga tggaatgaat atcggaaatt tatatgcagg tctagaacac 300 tcgcaatctc atgcagttcg tcactctctg ccttctgttc accatgtcca gccttttcaa 360 ggcccaccaa caacgagcac aacagtaact gtaccgcagc caccttcaat tcgtcctagg 420 gttcgagctc ggaggggaca agccacagat ccacatagca ttgctgagag gctgagaaga 480 gagaggatat ctgaaagaat aaaggctttg caggaactgg tacccagctg caataagaca 540 gatagggcag caatgcttga tgagattctg gactatgtga agttcttaag gcttcaagtt 600 aaggtactga gcatgagtag gctgggagga gccagtgcag tggcacaact tgttgctgac 660 attccattgc aatctgtgga gggggacagt ggtgaaagta gatccaacca gcgtatatgg 720 gataaatggt ccaatgtaga cacagaacaa gaggttgcta agctcatgga ggaagacgtc 780 ggtgcagcca tgcaatacct tcagtctaaa tcactctgca tcatgcccat atcacttgct 840 gcactcatct atcctactca gcaaccggat gaccagttac tggtcaagcc tgaagcggca 900 gccccatcat ag 912 <210> 59 <211> 687 <212> DNA <213> Solanum tuberosum <400> 59 atggcgtcaa tatcgattcc tttagcgtat aaatctcatt ctctccgtcg ttccccaatt 60 tatcgccctt cgcaacttca ctcacccatc caaatcaaat gttcagcttc caaggattgt 120 tccaacagtg aggaaagttc aactcagttt aagcaactta ggaatgttgc ttgtggattc 180 cttgctgttt gggcactttc ttctgtttct cccgttattg ctgctggtca gagattgcct 240 ccattatcaa cagatccaaa taggtgtgaa cgggcctttg ttggaagcac aataggtcaa 300 gccaatggtg tttatgataa accacttgat ctgcgctttt gtgattacac aaatgagaaa 360 actaatctta aggggaagtc acttgcagca gcactcatgt ctgatgcaaa atttgatggt 420 gcagacatga ctgaagtgat catgtctaag gcatatgctg ttggtgctag ttttaaggcc 480 atggactttt cgaatgctgt tctcgatcga gtgaactttg agaaagccaa cctccaggga 540 gcttcattta agaacactgt actatctgga tctaccttca atgatgcgca acttgatggt 600 gtagattttg aggacacaat cattggatac attgatctac agaagatatg cacaaataaa 660 actattaccg aagaaaggaa gagttaa 687 <210> 60 <211> 693 <212> DNA <213> Solanum tuberosum <400> 60 atggctgtga aagataaagc tgtgaaagga ggcaatgtga aggtgagtaa tggagttaag 60 gaggttcatt acagaggtgt aaggaagagg ccatggggtc gttacgctgc tgagattcgt 120 gacccgggta agaagagtcg ggtctggttg ggtacttttg atacggcgga ggaagcggcg 180 aaggcttacg ataccgccgc tagagacttt cgtggaccta aagctaagac gaatttcccc 240 tttccggcgg agattaatat taatagtaat aatcagagcc cttgtggaag cagcaccgtc 300 gagtcatcta gcggagaaac tgttgttcac gcgcctcata cgccacacgc gccgctagag 360 ttggatctca cgcgccgtct tggcgccgtt gctgaaggtg gacgtggagg tgtcggctac 420 ccggttttcc atcagcagcc gacggtggcg gtactgccga acggtcagcc ggttttgctg 480 tttgattcta tgtggagacc gggagttgtt agcaggccgt acaggttgtg ccggcgacga 540 tggagttcgc cggtgtcggt gccggagttg tttctggtgt gtcggattcg tcttccgttg 600 tggaagagaa tcgttacggg aaaaaaggac ttgatcttga tcttaacctt gcgccaccca 660 tggaagttta gtttgatgac gacgataacc taa 693 <210> 61 <211> 450 <212> DNA <213> Solanum tuberosum <400> 61 atggaattcg gccgcgcatt taatgccact tcaattttca ttgtttttct tcttgttact 60 ctagtttccg ctaagaagtc cggcgatgtt acggagttgc agatcggtgt gaagtttaag 120 ccaacatctt gcgaacttaa ggctcacaag ggcgatagag tctcagtaca ctacagtgga 180 aaacttacag atggaactgt atttgattcc agctatgaga ggaatgaccc cattgaattt 240 gaacttggaa gcggtcaagt gattaaaggt tgggatcaag gacttcttgg aatgtgtgtg 300 ggagagaagc gaaagttgaa aatccctgct aaacttggtt atagtgagag tggatctcca 360 ccaaagatcc caggtggtgc aacacttgtg ttcgacactg agctggttgc tgtgaatgga 420 aagaaaacaa caactgatag tgaactgtga 450 <210> 62 <211> 1032 <212> DNA <213> Solanum tuberosum <400> 62 atggctcatc tttcagatct tgtcaatctc aatctctctg attcctctga caaaatcatt 60 gctgaataca tatggattgg tggatcagga atggatgtaa ggagcaaagc caggactcta 120 tctggtcctg ttgatgatcc ttcaaagctt cccaaatgga attatgatgg ttctagcaca 180 ggtcaagctc ctggagaaga cagtgaagtg atcctatatc ctcaagcaat tttcaaggat 240 ccattcagga ggggcaacaa tatcttggtc atctgtgatt gttacacccc agctggtgaa 300 ccaattccaa caaacaagag gcacaatgct gctaaaatat ttagcaaccc tgatgttgtt 360 gttgaggaac catggtatgg tcttgagcaa gaatacacct tgctacaaaa ggaaattaat 420 tggcctcttg gatggcctat tggtggcttt cctggaccac agggaccata ctactgtgga 480 attggatctg gaaaggcttt tggacgcgat attgttgatg ctcattacaa ggcatgtatc 540 tatgccggga ttaacattag tggtatcaac ggagaagtga tgcctggaca gtgggaattt 600 caagttggac ctccagttgg cattgcatca gtgacgagtt gtgggcagct cgttacattc 660 tcgagaggat tacggagatt gctggagttg tcgtgtcact cgaccccaaa cctattccgg 720 gcgactggaa tggtgcagga gcgcatacaa attacagtac taagtctatg agatgaggag 780 ggtatgagtt atcagcgcta ttgagaagct tggacttagc acagagcaca ttgcagcata 840 tggtgaggca cgacgtcgtc tcactggaag accgaaacag ctgacatcac acgttcaaat 900 gggggtgttg caatctggtg catcattcct gtggaagaga ccgggaaaga aggcaggata 960 cttgaggaac agggaggcct gcatcgactt gaatcataca tcgtgacgtt tgatcggaac 1020 acctctcttt ga 1032 <210> 63 <211> 636 <212> DNA <213> Solanum tuberosum <400> 63 atggttaagc taagttgtgg tcctatttta ctagctctgg ttctttgcat cagcctaacc 60 tcagttgcca acgcacagca gtgcggaagg caaaggggag gagcgttatg tggcaacaac 120 ttatgttgca gccaattcgg gtggtgtgga tcgacacccg aatattgttc acctagccaa 180 ggttgccaga gccagtgtac tggaagtgga ccagatccag gccaaggggg cagcgcgcaa 240 aacgttcgtg caacatatca tatatataac ccacagaatg ttgggtggga tttgaatgct 300 gttagcgctt actgttctac ttgggatgct aataagcctt acgcctggcg gagtaagtat 360 ggttggactg ctttttgtgg tccagtcgga cctcgtggtc gtgactcgtg cggaaagtgc 420 ttaagggtga caaatacacg tacaggagct caaacgacgg tgagaatcgt ggatcaatgc 480 agcaacggcg gactagattt ggacattaac gtttttcaac aaatcgatac agatggagta 540 ggaaatcaac aaggacacct tattgtgaac taccaatttg ttaattgtgg tgacaatgtc 600 aatgttcctc tgctttctgt agttgacaaa gaatga 636 <210> 64 <211> 516 <212> DNA <213> Solanum tuberosum <400> 64 atggagcatg atgagacggg atgccaaccc cctccggaag gtcctatttt gtgtattaac 60 aactgtggct ttttcggaag tgctgcaaac atgaacatgt gctccaagtg ttacaaagac 120 atggtattga aacaagaaca agctaagctt gctgtgtcat caattgaaaa ccttgtcaat 180 ggatcatctg ccggagagaa gggaatggtt attgttgacc ccgtagatgt gcaacctgat 240 actattgaag cacagtctat agctttgcca tcatctcaaa cttcgagttc tagtgatatg 300 cctgatgtaa aggctaaggt gggtcctaac cgctgtggca cttgtaagaa aaaggttggc 360 atcactggat tcaaatgtcg ctgtggtaac ctttattgtg gagcacaccg ctactcggat 420 aaacatgact gcctgtttga ttaccgctct gctggccagg atgcaatagc aaaagccaac 480 cctgttgtta aggcagagaa gcttgacaag atctag 516 <210> 65 <211> 690 <212> DNA <213> Solanum tuberosum <400> 65 atgccgaagt cgaagcgtaa cagagctgtt accttatcta agacgaagaa gaagggtaag 60 gagcacaaag aaaacattgt caactcaata agggaatgcg cggaaaagta tagttcagcg 120 tacgtattta gtttcgagaa catgagaaat ctcaagttca aggagttcag agatcagctc 180 aagtcttcta gcagattttt cctgggatcg aacaaagtca tgcaggtagc actaggtcgg 240 tcagactctg atgagattcg tcccggcctc cataagattt ctaagcttgt acgcggtgat 300 tcaggtcttt gctttactaa cttatcgaag gaggaggtcc aaaggttgtt taacgagtat 360 gaagaacatg actttgcgag gactggcact acagcaactg aaaaggtgga gctacaagaa 420 ggtcctcttg atcagttcac acatgagatg gagccattct tgcggaaaca agggatgcca 480 gtgcggttga acagaggtgt cgtggagctc gtgtctgact ttgttgtatg tgaagaaggg 540 aggccattgt caccagaggc agctcggata ctgcgtctgc ttggtaagaa gatggctact 600 ttccggctcc acttaatatg caggtggtct cctgatgagt ttgaaatcta cagagaagga 660 ttggaagaat cagatattga atcctcatag 690 <210> 66 <211> 363 <212> DNA <213> Solanum tuberosum <400> 66 atggcaagga tcaaagttca cgagctgagg aacaagtcca agactgagct tttggctcag 60 ctgaaggacc tgaaagcaga actgtctctt ctccgcgttg ccaaagtcac tggcggtgcc 120 cccaacaagc tctcaaagat aaaggtagtt agattgtcaa tagcacaggt gttgacggtt 180 atatcgcaga agcagaagtt ggcacttagg gaagcataca agaacgagaa gtatttgcct 240 ctggatcttc gtcccaagaa aaccagagct attcgtaaac gtcttactaa gcaccaggca 300 tctttgaaga ccgaaagaga gaagatgtac tttccactta gaaagtatgc cattaaggtt 360 tga 363 <210> 67 <211> 585 <212> DNA <213> Solanum tuberosum <400> 67 atggcggcgg cgggattttt ctcaaatacc aaatactccg acagcttaac actcgtcgga 60 atttcaatct gcacagctat agtatgtgaa gcaatttcat atctcctcat ataccgcact 120 acttcataca aatccctcaa atctacaatc gataaagctt caaaaaagct cgaaactatg 180 aagactcagg aaccggttcc atctgtactc acgaagaagt cgaagacaaa gaagattgat 240 cgggtcgaaa cttccttgaa ggagtcaacc cgggatctct cccttttcaa attcaaatcc 300 ggattcgtcg tcgccgtagt gttgtttatg gttttcgggt ttcttaacag tctttttgaa 360 gggaaagttg ttgctaaatt gccgtttata ccctttcgtc ttgttcagaa gatgagtcat 420 agagggttac cgggtgatga catgacggat tgttcaatgg cgtttttgta cttcctttgt 480 tcgattagta tcaggacgaa tcttcagaag ttcttagggt tttcgcctcc tcgtggatct 540 gctggagctg ggttattccc aatgcctgat ccaaagacca attga 585 <210> 68 <211> 408 <212> DNA <213> Solanum tuberosum <400> 68 atggtgaaat tcctcaagcc aaacaaagcc gtagtggttc tccaaggcaa gtacgccggg 60 aggaaagcgg tgattgttcg agcgttcgac gatgggacac gtgataggcc atacggacat 120 tgtttggtcg ccggcgtagc taagtacccg aagaaagtgg tccgtaagga ctctgcgaaa 180 aagcaggcga ataaatcacg cgtcaagact ttcatcaagc ttgtcaatta caaccacatc 240 atgcctactc gttacacact ggatgttgat ttgaaggaag ccgttactgt cgattgcctt 300 cagtcacgtg ataagaaggt tactgctgct aaggaggcca aggctaagtt tgaagagcga 360 ttcaaaaccg ggaaaaatcg ctggttcttc actaagctta gattttga 408 <210> 69 <211> 654 <212> DNA <213> Solanum tuberosum <400> 69 atgggtgtta aagcaaggaa gaaagcaata acgaagaaat tgaaaaaggg agcaagcgat 60 ttttctgctt cttcagcttc taaagatgaa gcccgtgagt ttctgccatt agaaggtggg 120 ccagcgcaga agctcccaaa gactgaatta aaggaaaata aagcaacagt gttatatatc 180 gggagaatcc cacatggatt ttatgagaat gaaatggaag gtttctttaa gcaatttggt 240 accatcaaga ggcttagaat tgcaaggaat aaaaagacag gaaaatcaaa gcattttggc 300 ttcatagaat ttgagtctcc tgaggttgca aaagttgttg cggaaactat gcataactat 360 ctcttgtttg agcacttact gcaagttcag cttattcctc ctgaacatgt acatccaaga 420 ttgtggaaag gtgtgaaccg ctggtacaaa cccttggatt gggttaaaat tgaaaggaaa 480 cgtcaggaca aggaaagaac actggatgat cacaagaagt tggttgaagg aatattaaaa 540 cgtgacaaga agaggcgaaa gaggattgaa gctgctggaa ttgactatga gtgcccagaa 600 atggtgggtt caattcacgc tgcaccaaag aagatcagat ttgatgcaga ttag 654 <210> 70 <211> 237 <212> DNA <213> Solanum tuberosum <400> 70 atggctctat ccatgcgttt ctttgctact ttctttcttc tagctatgct tgtcatggct 60 actgagatgg gaccaatgag aattgcagag gcaagaaatt gcgaatcgtt gagccatcgt 120 ttcaagggac catgtacgag agatagcaat tgtgcttcgg tctgtgagac cgaaagattt 180 tccggtggca attgccatgg attccgtcgc cgttgcttct gcactaagcc atgctaa 237 <210> 71 <211> 384 <212> DNA <213> Solanum tuberosum <400> 71 atggtgttaa ttgaaaagct ttgggatgat gttatggctg gacctcaacc tgataaaggc 60 cttggcaaac ttagaaagtc cattactcta atgaaaactc atggcgggga aggaggagaa 120 ggagcaagca agagttacca aaggtctctg tctatgccgg gctcgggctt gtcgagcccg 180 gggacaccgg gaaccccggt gaccccaaac acttactcgc caacggcgcg taaagataat 240 gtgtggagga gcgtgttcaa tccggggagc aacattgcca ccaagagaat tggtgctgaa 300 gtctttgata aaccgtctca ccccaatgct cccaccgttt atgattggct ctacagtggg 360 gatacccgat ccaagcatca gtga 384 <210> 72 <211> 516 <212> DNA <213> Solanum tuberosum <400> 72 atggcaaatc caagggtttt ctttgacctt accatcggcg gtgcaccagc tggccgcgtg 60 gtgatggagc tcttcgccga taccactccc aaaaccgctg agaacttccg agctctctgt 120 accggtgaaa aaggtgttgg aaagatgggt aagcctttgc actacaaggg ctcaaccttc 180 caccgtgtga tcccagggtt catgtgtcaa ggaggtgatt tcaccgccgg aaacggtacc 240 ggaggagagt cgatctacgg cgccaaattc aaggacgaga acttcgtgaa gaagcacacc 300 ggcactggaa ttctctccat ggctaatgct ggacctggaa ccaacggttc ccagttcttc 360 atctgtaccg ctaagactga gtggctcgat ggaaagcacg tcgtgtttgg acaagttgtt 420 gaaggcttgg atgtgattaa gaaggccgag gctgttggat ctagttctgg aaggtgctcc 480 aagcctgttg tggttgctga ctgcggtcaa ctctag 516 <210> 73 <211> 441 <212> DNA <213> Solanum tuberosum <400> 73 atggcatcag ctctgttttc tgcaccaact ttccaaggtc tcagaccact caacaaaccc 60 actgactctt ccctcttcct caacaaaccc atcgccttca aacctatcac caaaaaaacc 120 cgaaatctcg ctgtgaaagc tgaactcaac ccatcaattg tcatcagttt gagcactggg 180 ctttcgcttt tccttggaag gttcgttttc tttaatttcc agagagaaaa tgtagcgaaa 240 caagtgccgt ctcaggatgg aatttctcac tttgaagctg gagatgttag ggctaaggaa 300 tatataagtt tgctaaaatc taatgatcca gttggattca atattgttga tgtgcttgct 360 tggggatcta ttggacatat tgttgcttat tatatccttg ctacttccag caatggatat 420 gatcccagtt tctttggctg a 441 <210> 74 <211> 375 <212> DNA <213> Solanum tuberosum <400> 74 atggtgttga ttgataaact ttgggatgaa gttatggctg gtcccagccc tgataaagga 60 ctaggcaaac tcagaaagag cctcactgtt caaactggtg ggggggaatc aggagaagga 120 tctagcaagt accagaggtc tctgtcgatg ccgacaagtc cgcctacgcc aggtacgccg 180 gtgacgccga caaacatttc gccgacggta cggaaagaga atgtatggag gagtgttttt 240 aacccaggga gcaatttagc taccaggaga attggtgctg aagtctttga taagccttct 300 catcctaacg ctcctactgt ttatgactgg ctctacagtg ggaacaccag atctaagcat 360 catgagaagt gctga 375 <210> 75 <211> 1117 <212> DNA <213> Solanum tuberosum <400> 75 gagcctctgt tctcaacggc ttgtttctca gaatggacat gccaaaagcc ccaaattcac 60 catacgtcct acactacaca tcttacgatt ccaaaactaa cggcgccggt gagaagtgca 120 ccctggaagt tgacgccgtt atcggcgcag acggcgcaaa ctcccgtgtc gcaaaatcca 180 ttaacgccgg cgactacgag tacgctattg cattccagga acgaattaga atttccgatg 240 ataaaatgaa gtactacgag aatttagctg aaatgtacgt cggtgatgac gtgtcccctg 300 atttctacgg ctgggttttc cccaaatgtg accacgttgc cgtcggcaca ggcactgtta 360 cccacaaagc agacattaag aaattccagc tagcaacaag actgagagcc gattccaaaa 420 tcaccggcgg gaaaatcatc cgagttgagg ctcacccaat tccagaacac ccaaggccta 480 gaagattgca agacagagtc gcattagttg gggatgcagc agggtacgtg accaaatgtt 540 caggtgaagg gatttacttc gcggcgaaga gtggacgtat gtgtgctgaa gcaattgttg 600 aagggtcaga aaatgggaag agaatggtag acgagagtga tttgaggaag tacttggaga 660 aatgggacaa gacttactgg cccacataca agtgcttgat atattgcaga agtattttac 720 aggtcgaatc cagcaaggaa gctttcgtgg agatgtgcgc tgatgagtat gtgcagagat 780 gacattcgac agctatttgt acagaaagtg gcaccaggga cccatgagac tgaagctgct 840 gtgataccat tggaagttgg tgaggctatg cactagagga aatgatagct catgtataag 900 aagaaggatg aattctaact agctaatatt tttaactgat gatgaagaat ttattctctt 960 atcgatttct cagtccaaaa aaaaaaaaat ggtgttcatg tacccgctct cggtcgatcc 1020 accgtacgtc atacattaca ttgatgttca ggccctggta taatatggct actgctcatc 1080 gtagacagag taccgatcgc tccaattctt aatggta 1117 <210> 76 <211> 1138 <212> DNA <213> Solanum tuberosum <400> 76 ggtactttga attggagctc tgttcaccaa atctagtgat ttctcgttag gttattatat 60 aaggcggctg ggatcggaac agttgagata gatttgccga tggcgcaaaa gatggctgat 120 tggtcggaat tgcaacttga tctgttggtt tcgataggta gatgtttgaa tctgattgaa 180 gactacctaa attttgggcg tgtctgcaaa tcctggcact ctgtggccac caaggccaat 240 tttaacaatg acctgtgtag agctccttgg ctgatgctag cggaggaaga ggataatgag 300 gttcggaaat tcttcagcct ctataatggc atgattttga acaagaggat tcccaaagct 360 agcagaaaac gctgtttgga gtctatgggt tggcttatca cggtaggaga agagggtgaa 420 attagtctgc tacatccctt ctctgatgtt cagatcgaat tgccccatca aaataccaca 480 gtagagtaca acaaacatca gactggcgca ctgacttgct ttattggcaa ggcagttttg 540 tcagctagtc cttcacacac atccgactac cttctcatgg tcattgatgg aggctcgagg 600 ttcctcagtt tttggagacc tggagatatt agatggacta gagttacatg ggaaggaatc 660 aattacagtt cttttgctga tttgatatat tttaatggcc aaatttatgc cgtggattat 720 tcgggtaatc tcctagtttg taatgttgct gatgttgtta gccctaaact tacaaaatgt 780 catatcatac catctcaacc tcatgaacat attccagagc acttatacat cctagaatca 840 ctagatcatt atttgtagct gtgcgatatg gtgttcactt tagcttgtcc agatgattgc 900 gacatgattc cattaacgct cgtccagacc ttatcacgtt gggtgagcta acatatggca 960 cagatatttc gagtttcccg gtgattagct gctcaaattg acgaaaccag attaggcacg 1020 cagctttttt ctagccgcta atgctccctc agtccagctt ccgatatcaa gaattaacat 1080 ccatttaatt accgagtagt tgtatgtaca tctacatgtg tccctcaatt ggtgacga 1138 <210> 77 <211> 859 <212> DNA <213> Solanum tuberosum <400> 77 gcagagtaca gggctgttaa gcctcatgag aggaacacta gatacgttga tgctgtcatg 60 acgattccaa aaggcacttt gttccccatg tgtggaatga acttggcatt tgaccgtgat 120 ctcattggac ctgcaatgta ctttggtctc atgggtgacg gtcagccaat tggtcgttac 180 gacgatatgt gggccggctg gtgtaccaag gtcatttgtg accatttggg actaggaatc 240 aagactggtc tgccctacat atggcacagc aaagccagca acccattcgt taacctcaaa 300 aaggagtaca acggtatctt ctggcaagag gagatcatac ccttcttcca ggctgcaact 360 ctttctaaag agtgcacaac cgtccaacaa tgctaccttg agctctcaaa gcaggtcaag 420 gaaaaacttt ccacgatcga tccctatttc accaagctag cagatgccat ggtcacatgg 480 atcgaagctt gggatgagct caaccctact ggtgacaact tggctaagct gtctatcgcc 540 gatggtccag caaagactaa aaagtagatg ctatttatag cttttgctca ttttcttcga 600 cgactaagac acttatgagt taactagaac tggattttac ttgtttatca atttacttgt 660 tttaatttta gttgaggatt taggtttcat atttgctgct tctactgtta tgtggagcag 720 cttttgattg ccttctctga gacatcatag gagtattttt attttattta tataaggctt 780 agttccaaac tttgagatct tctaatatca ataaaatatg gcttagtgct ttaaaaaaag 840 aaaaaaaaaa aaaaaaaaa 859 <210> 78 <211> 754 <212> DNA <213> Solanum tuberosum <400> 78 ggtacatgcg aagaaatgat gaaaagagct gtatttgcta gagaattggg cactccgatc 60 gtaatgcatg actacttaac ggggggattc accgcaaata ctgccttggc tcattattgc 120 cgagataatg gtctacttct tcacatccac cgtgcaatgc atgcggttat tgatagacag 180 aagaatcatg gtatgcactt ccgggtatta gcaaaagcgt tacgtatgtc tggtggagat 240 catattcact ctggtaccgt agtaggtaaa cttgaaggtg aaagagacat aactttgggc 300 tttgttgatt tactgcgtga tgatgttatt gaacaagata gaagtcgcgg tatttatttc 360 actcaagatt gggtctcttt accaggtgtt ctacctgtgg cttcaggagg tattcacgtt 420 tggcatatgc ctgctctgac cgagatcttt ggggatgatt ccgtactaca gttcggtgga 480 ggaactttag gacatccttg gggtaatgcg ccaggtgccg tagctaatcg agtatctcta 540 gaagcatgtg taaaagctcg taatgaagga cgtgatcttg ctcggaaaag gtaatgaaat 600 tattcgcgag gctgccaaat ggagcccgga actagctgct gcttgtgagg tatggaaaga 660 tatcgtattt aattttgcag caatggacgt ttttgataag taaaaacagt agacattagc 720 aaataaatta gcaaaaaaaa aaaaaaaaaa aaaa 754 <210> 79 <211> 1143 <212> DNA <213> Solanum tuberosum <400> 79 gaggggttct atctcttctt gtttgcttgc ttgttttctt tcttttcaag gttgctttta 60 aacccctttg agatattgaa tgagtaagaa caagacttat atggtgaaaa agttgctaaa 120 aaagtgaaga tctcaaattt atgtaggttt aatttctttt gagtttatat agtatagtat 180 atagtagcag agaggagcta gctatagctt aagggttttt ttttttttta agagaaaaga 240 agataagaag gaggaggaga agaagcttga gtagtatata gtacttttgc tttttcatta 300 ttcttcatct tgggattttc aagtgcctta aaaaaaagaa gcaaaacttc tcatattatt 360 attatttttg tttaattatg gagcctgcaa atctccatca acaatatcaa tatcatcaac 420 tccaattcca agatcaattt cctcttattg gtatttctcc aaattcttct tcatcttcta 480 ataattcttg ctatggagga gtttcaacaa caaacacttg gaccccatgt actactacta 540 ctactatctt gaacaatcat ggaagtggtc ttatgaattc atattctagt ggagacatca 600 tcaatactac aaattcatca tcaagtgatc cattaaacct agttaattct atgagtagta 660 ctactcaaga tttggaattt catcaatggg ctaataataa cattaagcaa gaaaattcat 720 atccaaaatt tactcaaatg ttgacaagtc catcaagtat agaagaagga agagaattga 780 gtgatatgaa tgctaagctt ttgcttggta cactttctaa tactgggctt caactttatc 840 atggagatat atataacaat aatcttctct atctcaaatc tagtagtatt agtactaata 900 gagggagatt agtcaatata tcccacaata atgtgtcatt tgatatatca gcaattcttg 960 tagctctttg gatatgatta cagcatgatt ccatcattta aagaattgga ggagtttagc 1020 gacaatgcct aacacaaatc atcgcatctc atgaagtccg tatagtccag attaagcatg 1080 atgatgacga gttgaggcac tggacttagt ctatgcaatc aagttacgca tgtcacctaa 1140 tga 1143 <210> 80 <211> 1157 <212> DNA <213> Solanum tuberosum <400> 80 gaagttatga ttgggtattc tgattcgggt aaggatgctg gccgactatc tgcagcttgg 60 caattgtaca aggctcaaga ggaacttgta aaggtggcaa aagaatttgg agtaaagcta 120 acaatgttcc atggccgagg tgggactgtt ggaaggggag ggggtcccac ccaccttgct 180 attttatcac aaccccccga cacaattctt ggctcacttc gtgtgactgt tcaaggtgaa 240 gttattgaac agtcatttgg ggaggaacac ctgtgcttta gaactcttca gcgtttcaca 300 gcagctacac ttgagcatgg aatgaacccc cctgtggctc caaagctaga gtggcgtgcc 360 ctattggatg agatgtctgt tattgctaca aaagagtatc ggtcttttgt tttccaggat 420 ccacgatttg ttgaatactt ccgccttgca accccagaat tagaatatgg tcgcatgaac 480 attggaagtc gcccggccaa aaggaagcca agtggtggga ttgaatctct tcgagctatt 540 ccatggatct ttgcatggac tcagacgagg tttcatctcc cagtctggct tggctttggg 600 gctgcattca agcacgttat tgagaaggat ataagaaatc tgcaaatgct caaggatatg 660 tacaacggat ggcctttctt tagagtgaca cttgacttac tcgaaatggt cttcgctaag 720 ggagaccctg gtattgctgc attgtacgat aagcttttag tatctgaaga tctgtggccg 780 cttggagagc gattgagatc caaatatgaa gaaacaaagg gctttctcct tcagttgccg 840 ggcacaagga tcttttggaa ggagatccat acttgaggca aaggctcaaa cttcgtgatt 900 cttacattac gaccctcaat gtaagccaag catacacatt gaaacgaatt cgcgacccca 960 actacaatgt gaaggtcagc cgcatctaga caggagatca tgggaatcag caaacagctg 1020 ctgaatagtg aagctgatct acgagcgagt atgcgctgct tgcagaccct tcatcctgga 1080 catgaagtat gctgctgcat gccagaacac ttgattagcc aagctattat gcaggtatta 1140 aatacgtgaa aactgtc 1157 <210> 81 <211> 1172 <212> DNA <213> Solanum tuberosum <400> 81 aaaaacgcaa atcgttttcc gttctggtaa caatgtcgaa acagacgtac aaagtcggat 60 tctttttccg gcggcagttt accatggctg ccgccgaagc tccggcagac attaagaacc 120 tatttaaacg ttattccgac gacagcggag ttatgagcgt ccagaatctt catcgtttct 180 taattgagat tcagaaggag aaaaatgcga gcttagataa cgctgaagcc attatcaata 240 atcacggcgg cgactccaag cagaaaggcc tccagctcga tggcttcttc aactgtctct 300 tttctgatgt taatcctcct ctcgatccta aacttgggat tcaccatgat atgaatgcac 360 ctctgtctca ttactacata tacacaggcc ataattcata tctaactggg aatcaactga 420 gtagtgattg cagtgatatc cccataatac aagctcttca gagaagtgtc agagtaattg 480 aattagatat ttggccaaat tcagacaaag atgatataga agttctacat ggaaggacac 540 tgactgctcc agtggcgcta atcaaatgtt tgaggtctat caaggagcat gccttttctg 600 catcggagta tcctgttgta ataacacttg aagatcatct cacgccagat cttcaggaaa 660 aagtggccga gatgatcacc cagacatttg gagacatgct gttttctcct tcagaaagtc 720 tgaaggagct tccttctccg gagtcactga gaaagcgtgt tatgatatca actaagccac 780 caaaagaata cttacaatcc aaggaagtta aaggaaaaag atgacacaaa gaaagaagct 840 gaggatgacg ttgatgagag agatgagatg agatgagatg atgagagatc caagtcagag 900 aggaggcagc gtcagataca gcgctatgca tcatgctgga aagggaaagg agactatctg 960 attgggtgag gtgactcata agtagacgct ggagtctcag gtgacagact gaaagctgta 1020 gaactcatca aagaaattat cagtcactca gcgaactggc tggaaaatac aagcatcgag 1080 tgactatcat taggatcctt gtggtggaat ctgactaagt gtaactaata gacaggtatt 1140 gagaaatcac tttgtgttga tagcactgaa gc 1172 <210> 82 <211> 1175 <212> DNA <213> Solanum tuberosum <400> 82 actttcagct ggtgcctggg ttgttacaac ttcagtatac ttaataaaca atcgggtaat 60 tggactaaaa gaagaggaaa ccgagaacct aaatcacaaa ttacacacac agagaagagt 120 tgaaccatga tgggaagatg cttatcagtt caatcatcaa acccaaccga gcttaaattt 180 caaactttga atggaattca acatctagct gaaagtcgtc gattcaaggc atggtttctg 240 gatcagtttg gtgtgcttca tgatgggaaa caaccctatc ctggtgccat ttcggccttg 300 gaaaagttgg caacttttgg tgcaaagatg gtaatcatta gtaattcttc aagaagagca 360 tctacaaccc tggaaaaatt gaggagcctt ggatttgacc cgtcactctt tattggtgcc 420 atcatgagtg gtgaattaac acatcagtat cttcaaacga gagaagacgc ttggtttgcc 480 tcaataggaa gatcgtgtat tcacatgact tggagtgaca ggggtgctat atctcttaaa 540 ggcctcggac tagaagttgt agaaaatgtc gaggaagctg attttatttt ggctcacgga 600 actgaagcct tggggctttc ttctggtgcc gcaattccta tgaatcttga tgaacttgag 660 aagatactgg atcggtgtgc tactaagaaa atccccatgg tagtagcaaa tcctgatttt 720 gtaactgttg aagctagagc tctgcgtgtt atgcctggaa cattggcagc cacgtatgaa 780 aaactggcgg tgaagtgaaa tggatgggta aacctgataa gataatatac aagtcagcta 840 tggaaatggc ttctgttgtg gatgcctccg attgtgttgc tataggggat tcccttcacc 900 atgatatcaa aggcgcaaat gtagctgaaa ttgcatcagc gttatcacag gcgatacatg 960 ctgctgaact tggactgcaa attcgagaag tcgcggatga tgatgctgta cacgctctgc 1020 tgaaaaacaa tgcaatatcc tacatatgtg ctcccttcat tacatgggtg aagaattact 1080 atcacttgta cctctttgga attcttggtg gtcacattgg tttatctgat tgttttcttg 1140 agcacactca catttatatc tctgtgtggg gggat 1175 <210> 83 <211> 1239 <212> DNA <213> Solanum tuberosum <400> 83 gatctccacc atcccaatca ctccttagcg gcggcgattt tatctcttct caatcagtct 60 ccgcctcaat tcccggcgcc gtcattttat tggacggaaa aagtgaaaac atgacggcga 120 ttcagacgtt tttccttcaa ggctcagttt caactctcct ttcttctcaa tcatcagtca 180 ctagctgtcg aaaagtttcc cttaaattca ccgtctcatg ttgctcggct acgtctcctc 240 tggccgtcgt gaacggtaat gtggagaaga gaccttccga tagaaccgaa gttcgcatcg 300 gtttgcctag caaaggccga atggctgctg attctctcga tcttctcaag gattgccagc 360 tatcagttag gcaggtgtat ccgcggcagt acgttgcaga aattccacag atacccaatc 420 tggaagtgtg gtttcaacgg ccaaaagaca ttgtgcggaa gttggtagct ggagatttag 480 accttggtat tgttggtctc gacacagttt gtgaatatgg gcagggggat ggagatctca 540 tcattgtcca tgatgccctt gagtatggtg attgtcgttt atctcttgct ataccaaaat 600 atgggatttt tgagaatgtg aactcaatga aggaattagc agagatgcca cagtggacac 660 ctgagaggcc actgagagtt gcaacaggat tcacatatat gggtcctaaa tttatgaaag 720 aaaatgggtt gaagcatgtc actttttcaa ctgctgatgg agccctggaa gcagctccag 780 cgatgggaat agctgatgct attgtggatc tagtgagcag cggaaccaca ctgagagaaa 840 acaatctgaa ggaaatagaa aggtggagtc atcgtgcaaa gtcaggcggt ccttgtcgct 900 ggcagaaagt ccctaatgca gcgaaaaggt gcacttgata taacacatga aatgctagaa 960 agactggagg caccattctg gaggggctga aggtcaattt acggtaactg ctaatatgcc 1020 gaggaagcag tgcaaaggaa gtccctgaac gaatattgga gccaaacgtc actatcctgg 1080 gattgcaggt accaccatta agttcagtct ttgccaccgg agtgaacgag ggttcccgcc 1140 tgaatacttg gccattgttc atatgtgtta acccaagaaa gacaccttac acagccgtgt 1200 tccagcgacg cttatatggt cggcatgatg atgatgtat 1239 <210> 84 <211> 1097 <212> DNA <213> Solanum tuberosum <400> 84 gctcctcctc ttctctctct attactatat aaaacctata gatacatata ttttctctat 60 tcatcgtcgc catggcagtt aacggtagtg ctccttcttc tggtttgatc gtcagtttcg 120 gtgagatgtt gatcgatttc gttccgacag tctccggcgt atcccttgcc gaggctcccg 180 gatttttgaa agctccgggc ggtgcaccgg cgaacgtcgc catcgcggtg acaaggctcg 240 gagggaagtc ggcgttcgtc gggaaactcg gcgacgatga gtttggtcac atgctcgccg 300 ggattctgaa aacgaacggc gtacaagccg acggaatcaa ttttgacaag ggcgccagga 360 cggctttggc gttcgtgact ctacgcgccg acggagagcg tgagtttatg ttttacagaa 420 atcccagtgc tgatatgttg ctcacgcccg atgagttgaa tcttgatctt attagatctg 480 ctaaggtgtt ccactacgga tcaattagtt tgatcgtgga gccatgtaga tcagcacatt 540 tgaaggcaat ggaagtagca aaggaggcag gggcattgct ctcttatgac ccaaaccttc 600 gtttgccgct gtggtcttcg gaagaagagg ccaggaagca aatcaagagc atctggaact 660 atgctgatgt gatcaaggtc agcgatgtgg aactcgaatt cctaactgga agcaacaaga 720 ttgatgatga atccgccatg tccttgtggc atcctaattt gaagctcctc ttggtcactc 780 ttggtgaaaa gggttgcaat tactacacca agaaattcca tggaagcgtt ggaggattcc 840 acgtgagact gttgacacca ctgagctggt gattcttttg tgtgctctct acagatgtga 900 tgatcagcat tctgagatga gcagtgagag tactagtttc atgtgcatgt gagcatcaca 960 cacagaagga gcatccagct ctgctactga atctgagcct cactgctcag gaaggacaga 1020 acatcaggta actttcattt ttcatccaat cccccttaga ctactcacgt ggagctcata 1080 gtgatagttt tgggacg 1097 <210> 85 <211> 1227 <212> DNA <213> Solanum tuberosum <400> 85 attccggtca caactatgga gtccaaagtc ctcccaattg ctccggcgcc gttaccgaca 60 ctttctgcct ttccgactac ctccgatgaa gccacggaat ggaatattca aagtccggtt 120 gcaaagtcgc cggcgccacc aacaacgcca tcgacgccgt catcctttca agatatacgg 180 cgatggagac cggcggcgca acgaaacctg agaaaccagt ggtcgaagtt gtcttctctt 240 cggacgcaat ggctttcttt atcctccact gctcgatcat acgccactta tgtagtaaat 300 tcttatctct cccaaaggta tatggatgca atggagttag gtgtattgac agagatgcct 360 gatatacgta aaaagcatgt aggaaattgt tcaagcagca ggagactaat aggagtaagc 420 ttttgtcatc ttacaaagat atggtggttg ttgtaactca gatggtcaat gtttctaaat 480 caatgaggtg ttatcacaaa ggagcaagtg gcaccccact tactgagttt tcttgcttcc 540 ctggaaacct gaatgacact ggtgattgtg atgggattcc tgttctcacc tttttgtcga 600 tctttgattt cgagaaactg gcagaggagc tggtccagat gtttgtgtca gaaacaaatc 660 taaagcggtt gctagtgatg gaaatatgct ccattggttc cgagaatttt tcacaagttg 720 acaggctgaa atggagtgat catttctatg ttggagagtt tgatcatcta cgcatatgtg 780 attcaaactc aaatgaggtt cttaagcaac ttgtgcccaa ggtagagagc tgcaattctc 840 agtctaccac catgcaatca agtagtcaac tagagaggac gttctccagg tttacttaac 900 aacctgctcg cagaagtcaa tgtcgataga ttcaggataa gtgattacta gctatggtag 960 gtgaggagat gcatgttacc atatcatgaa actcactaca cagaataggg cagaatggat 1020 acaaaggatt cacatggatg atcccaacta aattgagggt ggagggtgta taaaactgac 1080 ttaaattggg atttaaatgg tgaggaacta ttcacttaca aaacaagaac tactggcgtg 1140 tttcaggcaa tccgccaata tgggttgtct aatggcgctc ttgtgtgtgt ctagagacac 1200 acacacacgc gcggggagtg atgaaag 1227 <210> 86 <211> 1168 <212> DNA <213> Solanum tuberosum <400> 86 gatctagaaa aagagaaagt agatacaaaa tgcagaaagt ttgagacaaa aactgcaaaa 60 ttccatcaat taaatctcaa tttaagttgg cataataatc ttggttagcg tacctccaac 120 aacaactacg cccaagtccc aaacaagttg tgatattgtt agtacctccg aaaggaaata 180 atcatctcag cagtgaatta gtagagtgtc tagttgcttt ctgacacatg ggaagtattg 240 aggatgggaa gtcttctacg cctgagaaat cattttcgac tgcacctgat caaagcaatt 300 ttcatgtgta tcctgattgg gctgcgatgc aggcatatta tggtcatcga gtagctgtac 360 ctccatatgt taattctgct gttgcacctg gtcatgctcc tcatccttac atgtggggcc 420 cgctaccgcc tatgatgcca ccttttggca caccatatgc aggagtctat gcacatggtg 480 gtgtttatcc gcaccctgga gttcctattg tgtcccgtcc tcaggcacat ggaatgacat 540 catctcctgc tgttagccaa accatgaata ctgcttcttt gagtatggat gcttctgcta 600 agtcttcggg gaactctgat caaggcttga cgagtcagtc aaaagggttt aatgggcttg 660 gaatgtcaat tggaaatttc agtgttgaca atggtgatgg cactgaccat ggaccttctc 720 agagtgggca aactgaaggt tcaagtgatg gaagtaacat acacacagca gaggttggtg 780 agcacaggaa gaaaagaagc cgcgaggcaa ctcctaataa ctctggtgac agaaaagtca 840 aagactgtgc agtccacaac gtactaggga agtaaatggg gactcagaga aggaatcggt 900 gtagctgtat cctgtatata gcagaaaatt agcgggacac attttctcca tcatgactac 960 tactcgctag taaatctagt cactcacgat cagtctgcag tgcggcgagt ctgtacgatg 1020 acgtgagctg agcgggagaa gagagcagtc ttatcgagat ctgcaggaat caggtggaaa 1080 acgccgagct gagatggcag tgcggatcaa cttactcgat tgactagaat cgataccatc 1140 gcgaactttg gaacataagg ttgaaagt 1168 <210> 87 <211> 1172 <212> DNA <213> Solanum tuberosum <400> 87 gccttcatca cttttgctgt gattcttctt cagctgtgca gcatatcgcc taactcttct 60 ctaagtttct taacctctca agatgggtaa agagaagatt cacatcagca ttgtggtcat 120 tggccatgtc gactctggaa agtcgactac cactggtcac ttgatctaca agcttggtgg 180 aattgacaag cgtgttattg agaggttcga gaaggaagct gctgagatga acaagaggtc 240 attcaagtat gcctgggtgc ttgacaaact taaggccgaa cgtgagcgtg gtatcaccat 300 tgatattgct ttgtggaagt ttgagaccac taagtactac tgcactgtga ttgatgcccc 360 cggacacagg gacttcatca agaacatgat cactggtacc tcccaggctg actgtgctgt 420 cctcattatt gactccacca ctggtggttt tgaagctggt atctctaaag atggtcagac 480 acgtgaacat gcattgcttg cttttaccct tggtgtcaaa caaatgattt gctgctgtaa 540 caagatggat gctaccaccc ccaaatactc caaggctagg tatgatgaaa tcgtgaagga 600 agtttcttcc tatctcaaga aggtcggtta caaccctgac aaaatcccat tcgtccccat 660 ctccggtttt gaaggagata acatgattga gaggtctacc aaccttgact ggtacaaggg 720 cccaaccctc cttgatgctc tcgaccagat taatgagccc aagaggccaa cagacaaacc 780 cctccgtctt ccacttcagg atgtttacaa gattggtggt attggaactg tccctgttgg 840 tcgtgttgag actggtgtga tcaagcctgg tatggttgtg acctttggcc ctactggttt 900 gacaactgaa gtcaagtctg ttgagatgca cccacgaagc tcttctggag gcactccctg 960 tgacaatgtt ggggttcaat gtagaatgtt gctgttaaga tctgaagcgt gattttgttg 1020 ctcaactcca gatgacccag ctaggggcag cagcttcact gccagtcatc aatcatgaca 1080 tctgccaatt gaaacggaat gctcaatggc tgactgtcac tcccacattg cctgttaagg 1140 ttgtctgtga aatctctgtg acacagaaaa at 1172 <210> 88 <211> 1225 <212> DNA <213> Solanum tuberosum <400> 88 cgggttgagc tcactaaaca gtatcatcaa gctcttgatg cttccgttgc tcggaatttc 60 cctgataatg gttgcattgc ttgcatgagc cacagtactg atgcgcttta ctgttcaaag 120 cagacggcag ttgtgagagc atcagatgac ttctatccaa gagatccagc ttcacacacc 180 attcacattg cttgtgtggc atacaacagt gtgttccttg gagaaattat gcagcctgat 240 tgggacatgt tccactccct tcatccagca gccgagtagc atggctcggc gagggcgttg 300 agtggtgaac ctgtctatgt tatggatgca cctggcaagc acaactttga tgttctgagg 360 aagcttgttc ttcccgatgg ctcaattctt cgtgctcgtt tgcctggtcg acctacaaag 420 gactcccttt tcactgatcc ttctcgtgat ggtgttagcc ttttgaagat atggaacatg 480 aacaaataca atggtgttct tggaatatac aactgccaag gtgcagcgtg gagcacggct 540 gagaggaaga ccacgttcca caaaaccaac tcagaggcga taacaggcta tataaggggc 600 cgtgatgtcc atttcatatc tgaagctgcc ttggacccca actggagtgg agacacagtt 660 ctttactctc atggaagtgc tgagcttgtt gttctcccgt ataatgcagc aatgcctgtt 720 tcttttaaga tccttgagca cgagacatac actgtcacgc ctattaaggt cttggcacct 780 ggctccagct ttgcaccttt ggggctcatt gacatgtaca atgctggtgg ggcaattgaa 840 ggactaaaat atgaggtgaa ggcaggcgca gaattgtctg aactcgagct ggatatcaag 900 gtgaaggaat cttgtggctg aagataaaat tgaggacttg agcacagaag cagttgcagt 960 agtgtcgatg gaagtaagtg gatgtggcgg atttgtgtta ctcgtctgtc agcgaggatt 1020 atgggggggg gtgttgtgtt gtttttttgc ccaaactcgc gagatcgggg tttgtaaact 1080 tggaactctg aggcatggcc tccgagcgaa atcggaagtg gcatatccat tggaagtgtg 1140 aagattagaa ttacgagggg agaccatgtg ccctgactac ataagagcgc gaatctttgc 1200 ggactcatac agagagcgag tatag 1225 <210> 89 <211> 1112 <212> DNA <213> Solanum tuberosum <400> 89 atcccaaagt atatattttg ttctactaat actacatttt ttttcataca aaattatcac 60 caaaaaacat catggctgaa gagaagaaac atcactttgg tggtctcttc caccaccaca 120 agaacgagga ggaagacacc cccgttgaga aaactacata tgaagaaaca acctatggag 180 agagtgaaaa aactagaacc tatggtgaga aaactagtta tggtgatgat acatatggcg 240 aaaaaaacta ccacattcgg agatgacaac aaatatggtg agaaaactag ttatggtgat 300 gatacatatg gtgaaaaacc tacctcctac ggaggagata atacatatgg tgagaaaaca 360 agttatggtg agggagatga caacaagtat ggtgagaaaa ctagttatgg tgagggagat 420 gacaacaagt atggagagaa aactagttat ggagatagcg gatacggaga gaaacctagt 480 tatggtgggg ggagatgaca acaaatatgg tgagaaaact agttatggta atgaggaagg 540 tggctatggt ggaggagttt ggtgagacta ctaattatga agagaatgaa agtgaaacca 600 aaacttctga ggattataag gaagagaaga aacaccacaa gcatcttgaa gaaattgggg 660 gactcggagc tgttgctgct ggtgcctttg ctttgcatga gaagcacaag gcagagaaag 720 acccagagaa tgcacacaag cacaagatag aggatggaat agcagcagct gctgcaattg 780 gtgctggtgg atttgcattc catgagcatc atgataagaa agaagcacag gagaagagga 840 aggacgctga gggtaatagg aagcatcatt tcttcactaa ctctcctatc ataatatata 900 gacttacgtc tgctttgtgc gtgctgatca atctatctta gtatactcgg catgcaccta 960 actagttagt atgtgtgtac atcttcttgc gattagatgt aacgatgctt ggagatagac 1020 agttcagtca ctgacgttgc aatacgctgc atgcactgtg aaaactccat acgatcaagt 1080 tcttattctc gagagagaaa cccgagacag ag 1112 <210> 90 <211> 1149 <212> DNA <213> Solanum tuberosum <400> 90 gcttctcaca cactctgtaa accctagctg tacacccaat acccacaatc atggcggaga 60 gaggaggagg ggagagaggt ggattcggcc gtggatttgg aggacgtggt gggagaggtg 120 gagaccgcgg tggacggggc cgtggaggcc gtcgtccacg tcgtgaaaca gaggaggaga 180 aatgggttcc ggtcacaaag ctaggaaggc tcgttaagga tggcaaaatc cgctcacttg 240 agcaaatcta ccttcactca ctcccaatca aggagtttca aatcatcgat actctcatcg 300 gtccatctct caaggatgag gtgatgaaaa tcatgccggt tcagaagcag acccgtgccg 360 gacagagaac caggttcaag gcgtttgttg ttgtaggtga cgggaatggt cacgttggtt 420 tgggtgttaa gtgctcgaag gaagtcgcta ctgctattcg tggaggaatt atattggcta 480 agctatcagt gattccagtg aggagaggtt actggggtaa caagattggt aagccacaca 540 ctgtgccttg caaggttacg gggaaatgtg ggtctgttac tgtgaggatg gtgcctgctc 600 ctcgtggtgc tggtattgtt gctgctcgtg tccccaagaa ggtccttcag tttgctggta 660 ttgaagatgt cttcacatct tctcgtggat ccaccaaaac ccttggcaac ttcgttaagg 720 ctacctttga ttgtttaatg aagacttacg gattcctgac cccagacttc tggaaagaga 780 ctcgtttcac caaatctcct ttccaagagt acttttgata tcttggcaaa accagcgaat 840 aaagttattg tctacgccac cgaagagcag cagcgcctgg agagggttga gcttgattca 900 ttagatgtgt atgagatctg tgtttggtct agtagtctct ctctctgata cgtagtattc 960 taaatttgtt ctgttttctg aggatgttta tccgcagact gcaaacataa tgtccctgtt 1020 ttggattgtt tcacaatacg tatggatttc taaaaaagaa taaagatggt gttccatcgt 1080 acccgttcct cgttcgatca ccggtcgtct cataccgatc catggatttc tgaaggcgcg 1140 acacagggt 1149 <210> 91 <211> 702 <212> DNA <213> Solanum tuberosum <400> 91 taggtgcaag gtaccaggtc attgcacaga tgagggtaca aaaatagtag taacagatca 60 tggtgaaggt gaccatacag attttatact aagtgtacgt gcttattcag acatggctac 120 ctcaggaatg gctaatcatt tgttggctta tggtgttgtt gacgttgaat atcgaagggt 180 accttgtacg tactatggtt acaatttaat gattaaagtt catgaacata gcagattttg 240 taactatttg gctattgtac caatttatca aagtggtgca tttgacattg aagctgtcga 300 ggtttggcag gctgattgca aggaatggag gggtatgaga aaggcctatg gagcagtatg 360 ggacatgcca aatccaccaa aaggatcact cactttcaga gttcaagtta ttcttaatgg 420 ggaggcagct aaatgggtgc aattggctga tgttttacct gatgaatgga aggctggcat 480 tgcttatgac acatacctta tgctagacta aatagtaata ttatgcttat ttactatgtt 540 gtataatgtc atgtggttag caaagtgttt tatttgaaga tttatcaaat aaactataga 600 agattaagta aatttgtaat gttgtagggt atgcattgtg gtgaattgtg tgtttgatat 660 aaataaaaag tcatacagtt tacaaaaaaa aaaaaaaaaa aa 702 <210> 92 <211> 1161 <212> DNA <213> Solanum tuberosum <400> 92 gaacctcata tcaactacat ttctattccc ccctttcttc taaacttgac atccctcttt 60 cttttgtggc tattcttgaa aaaaaataat tctttaatct atggctactc tcacaaattt 120 tttgctcaaa ccctctccaa atctagcttc aattactaaa attaaccctt cactttactc 180 caatttccct tttgagaaat ccaaacaatc aattttcaag aacctcaaaa caaacaagca 240 cttattgatt acaaaggcaa cagcagcacc tgatgttgag aaaaaagtgg ccaaaagtga 300 aagagtacaa aaggttaatg gtatggagga attggatgaa gcattaaaaa aggcgaaaaa 360 tcgactagtt gtggttgaat ttgcaggcaa agatagtgaa cgttccaaaa atatttatcc 420 ttttatggtg aatttaagta agacgtgtaa cgatgtcgat ttcttgttgg tgatcggaga 480 tgaaacggag aagactaagg cactctgccg gagggaaaag attgacaagg taccacattc 540 aacttctaca aaagcatgga gaagatccac gaggaagaag gcattggccc ggaccaacta 600 gccggtgatg tactctacta cggtgacagc cactccgaag tagtgcagct ccacagcaga 660 gaagacgttg aaaaggtaat ccaagatcac aaaatcgata aaaagttaat agtcctcgat 720 gttggattgt agcattgtgg accatgtgtg aaagtttatc caacggtgat caagctatcg 780 aagcagatgg ctgatacagt cgtgttcgcg agaatgaatg gcgatgagaa tgatagttgt 840 atgcagtttt gaaggacatg gaggttattg aagtgctaca ttttgttata agagatggtg 900 aaattgtgaa aggggtttgt tgtttcctgg ggaaaggcgc acttattgga gacattctta 960 gcatacagga gttacgggta cttattaagc attatatact cattgggact tggaataggg 1020 gtgactgatc tcgggaccag ctagtggaaa acacacagtg agtggttcga ataagttggg 1080 tggttcatcg cactagtcgc aacgagtgcc atgccgaata gcttgagcaa ccgcttatac 1140 ctaccaggaa ccggggtcat g 1161 <210> 93 <211> 713 <212> DNA <213> Solanum tuberosum <400> 93 atctcataca cataaacgca caattcattc ttcttctcta gaaaatcatg aattttctct 60 catctttggc taagagcgcc ggagccggcg atgagaaaaa agccggcggc gaggaatctg 120 gttccacaac agagcttttc gccagtgcta aagtgttagc agatgccgct caaagtcaat 180 tcaacaaaga ttccggcaaa gttgataaca aaaaagtagc tgaagcagct gccgatgtcc 240 tcgacgctgc acagaaatat ggaaaattag acgaaactca aggaattgga cagtacgttg 300 agaaagccga aacttatctc caccagtacg gtggcgataa ggctacggcc gccgctccgg 360 ttgctgaaga agcaaaagca cctgctcctg caccggaatc ggctccggca tcggaaccag 420 caccggcgcc ggttgttgaa gaagataagg aggaggaaaa aaagagtgga ggagctgaat 480 atcttaagat ggctggtgat ttcttaagca agaagtaatt agagtctaat taactttttt 540 taattatggt ctgtgctttt aattatggct gatgttatat ttcagttttt ttttcttaat 600 ttgccactgt ccactaccgg cgccggcgcc ggagttgggt ttatgtaata aaatatgtat 660 gtaagttgtg tgtattcaag tgtgatttta ccgcaaaaaa aaaaaaaaaa aaa 713 <210> 94 <211> 747 <212> DNA <213> Solanum tuberosum <400> 94 tacgccgctg caccaccgcc gagttccgtt agtttccggc agcggccgtt tctgtttccg 60 ccgcttactc cactgcggag aggacttcta ctactactac tagtagtagc tctacaatag 120 cctcacatac atcgttatac gaagttttat ggattcaaat aggagctaat tctcatgaaa 180 ttaaatctgc ttaccggaaa ttagccagaa ttttgcatcc ggatgttcag aattcgtcgg 240 cggaggactt tattagagtt caatcagcgt atgcttctct ttccgacccg gaaaaacgtg 300 ctaattatga tcggaaacta tttgggaata gaattgcaag gtctgttgat ttctcaccgg 360 cgggagctcg tagccattat actgttcgcc ggaaatggga aaccgatcag tgttggtaga 420 ttttttcgaa atctcccgcc cgattagtca taaaatgtag attttagctc tcctaatcga 480 gcgggactta atttctgatc ataactcaaa tctgaaatct cggattaaga ataataagat 540 cagttatttg tacagtacat gtaaatgtga ctttatgtat atattgcttt aaaattttgt 600 tatgttattt gtgtggaaga taaataatca gacgaagttt ttgaagtctg aaagttcatc 660 aaaattgtcc aatcaacatt tgtatattat tagaattcac aatataatat accttttttt 720 ataaccacaa aaaaaaaaaa aaaaaaa 747 <210> 95 <211> 1168 <212> DNA <213> Solanum tuberosum <400> 95 aaagggcggt ggtatatccc attaattagt agtattagta ttatattagt ccattgtttt 60 ccattttcca ctacctcatc agttagcttt gtgctttgaa agttccattt tcaagtttcc 120 aatctgagaa taaactataa tggcagaatc atcgccggta cgttttggca ttttaggttg 180 cgctaacatt gctcgtaaag tatcacgcgc catcgcactt gctccaaacg ccactatctc 240 tgccgtcggc agccgttcga tcgaaaaagc gacggcgtac gccaaggaaa acgggtatcc 300 gtcaacgacg aagatttacg gcagttatga ggcggttctg gatgatccgg aagtggatgc 360 cgtttatgtt cctcttccga cgagcctgca tgtgaagtgg gccgtgttgg cggcccagaa 420 gaagaagcat gttttgctgg aaaagcctgt tgcgttaaac gtgaaggagc tggatgtgat 480 actggaggag tgtgaatcga atggggtgca gtacatggat gctactatgt ggatgcatca 540 tccgcgtact gctaagatga aagagttcct ctctgattct cagcgttttg gacaactcaa 600 atcggtacat agcacgtttg cttatcttgg tgacacggaa tttcttgaga ataacattcg 660 cgtgaagtcc gatcttgatg ctcttggtgc tctaggtgat gctggttggt acagtattcg 720 ggcaatcttg tggagtagtg attatgaatt gccaaaaaca gtgacagctc tgcgtgatcc 780 agaatttaat gaagctggag ttatcctatc ttgtggcgct tctttgagtt ggaaagacgg 840 aaggtagcaa ctttttattg ttcgttttta gccatatggc catggatatc gttgctagtg 900 gatccaaagg aatctgcggt ccatgacttt gtattccatt tcagaatgtg ctcatttacc 960 actgtggaag gttctttggt ttggtgactg ctacatcgat catcctgcac cagtgagcat 1020 caggtcaata cgatcttcct cagagcttgt tgatggtaaa gagattcacc tgggtgggga 1080 ggacttacaa gggaggtcta cctggaaagt tgccagattg agccacctgg tgtgggagtg 1140 ctcagaccat tgaagttccg aaccgtga 1168 <210> 96 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> primer 1 <400> 96 ttgataccac tgcttagggc gagcttaata ttccct 36 <210> 97 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> primer 2 <400> 97 tctcatcgta ccccgctcct cggtctcgat tctacg 36 <210> 98 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> sequencing primer <400> 98 aatatacctc tatactttaa cgtc 24

Claims

고온 스트레스 저항성 유전자를 포함하는 고온 스트레스 저항성 형질전환체 제조용 발현벡터로서, 상기 유전자는 서열번호 1 내지 서열번호 95로 이루어진 군으로부터 선택되는 유전자인 발현벡터.
청구항 1에 있어서,
상기 유전자는 감자(Solanum tuberosum) 유래인 발현벡터.
청구항 1의 발현벡터를 숙주 세포에 도입하여 제조한 고온 스트레스 저항성 형질전환체.
청구항 3에 있어서,
상기 형질전환체는 식물체인 형질전환체.
청구항 4에 있어서,
상기 식물체는 감자, 양파, 가지, 담배, 당근, 오이, 올리브, 고구마, 배추, 양배추, 무, 수박, 참외, 오이, 호박, 딸기, 대두, 녹두, 강낭콩, 상추, 고추, 토마토, 브로콜리, 해바라기, 갓, 잔디, 애기장대, 유채, 및 갓으로 이루어진 군으로부터 선택되는 것인 형질전환체.
청구항 1의 발현벡터를 제조하는 단계;
상기 발현벡터를 숙주 식물세포에 도입하여 형질전환 식물세포를 제조하는 단계; 및
상기 형질전환 식물세포를 조직 배양하여 재분화하는 단계를 포함하는 고온 스트레스 저항성을 갖는 식물체의 제조 방법.
청구항 6에 있어서,
상기 발현벡터의 숙주 식물세포로의 도입은 아그로박테리움-매개에 의한 방법, 유전자총, 실리콘 탄화물 위스커법, 초음파 처리법, 전기천공법 및 PEG 침전법으로 이루어진 군으로부터 선택되는 것인 제조 방법.
청구항 1의 발현벡터를 식물체에 도입하는 단계를 포함하는 식물체의 고온 스트레스 저항성을 증가시키는 방법.