KR20230153145A - 식물체의 재분화 효율 조절에 관여하는 애기장대 유래 유전자 및 이의 용도 - Google Patents
식물체의 재분화 효율 조절에 관여하는 애기장대 유래 유전자 및 이의 용도 Download PDFInfo
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Abstract
본 발명은 식물체의 재분화 효율 조절에 관여하는 애기장대 유래 LUX, IDD7/11, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 및 COP1 유전자 및 이의 용도에 관한 것으로, 상기 애기장대 유래 유전자는 식물체의 재분화 효율을 효과적으로 개선할 수 있으므로 작물 형질전환 효율 증진에 기여할 수 있을 것이다.
Description
본 발명은 식물체의 재분화 효율 조절에 관여하는 애기장대 유래 LUX, IDD7/11, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 및 COP1 유전자 및 이의 용도에 관한 것이다.
식물의 재분화 (regeneration)는 작물의 게놈 (genome) 편집, 형질전환 등 유전공학적 응용에 필수적인 기술로서, 낮은 재분화 효율은 게놈 편집의 주요한 허들로 알려져 있다. 고부가가치의 작물을 농생명공학적으로 육종하기 위해서는 게놈 편집 또는 형질전환후 조직의 탈분화 (캘러스 형성, callus formation)와 지상부 재분화 (shoot regeneration)가 순차적으로 수행된다. 이때 식물 재분화 과정의 효율이 매우 낮아 주요 작물의 형질개량이 매우 제한적으로 이루어지고 있는 상황이다.
최근 크리스퍼 (CRISPR) 기술의 발전과 함께 정밀 육종의 수요와 시장이 폭발적으로 성장하고 있으며, 식물 재분화 기술은 해당 시장에서 필수적으로 요구되는 바, 혁신적인 문제점 개선을 위해 전세계적으로 노력이 집중되고 있다. 일반적으로, 식물의 재분화 효율 증진을 위해 재분화 유도배지에 다양한 호르몬과 첨가물을 배합하는 방식으로 재분화 증진을 도모하였으나, 뚜렷한 한계에 직면하고 있다.
식물의 낮은 재분화 효율은 유전적 장벽 때문인 것으로 알려져 있는데, 식물의 유전적인 요인에 의해 재분화 효율이 이미 결정되는 상황이기 때문에 재분화 과정을 방해하는 유전자의 제거가 재분화 증진을 가능하게 하는 핵심기술로 알려지고 있다. 따라서 재분화 효율 조절에 관여하는 유전자를 확보하고 해당 유전자의 발현 조절을 통해 작물 재분화 효율을 증진하는 방법의 연구가 필요하다.
본 발명에서는 재분화에 관련된 중요 유전자를 선발하여 이 유전자의 발현을 조절하는 방식으로 재분화 효율을 개선할 수 있는 방법을 제안하고자 하였다.
한국등록특허 제1898392호에는 '애기장대 유래 ATX4 유전자를 이용한 식물체의 재분화 효율 증가 방법 및 그에 따른 식물체'가 개시되어 있고, 한국등록특허 제2125714호에는 '식물체의 재분화 효율을 조절하는 애기장대 유래 DEMETER 유전자 및 이의 용도'가 개시되어 있으나, 본 발명의 '식물체의 재분화 효율 조절에 관여하는 애기장대 유래 유전자 및 이의 용도'에 대해서는 기재된 바가 없다.
본 발명은 상기와 같은 요구에 의해 도출된 것으로서, 본 발명에서는 탈분화 및 재분화 과정에서 발현 변화가 동반되는 주요 유전자들을 대량으로 스크리닝하였고, 주요 녹아웃 돌연변이체를 선발하여 재분화 과정에서 어떠한 영향이 있는지 확인하였다. 그 결과, LUX, IDD7/11 (IDD7 / IDD11), SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 및 COP1 유전자는 모두 재분화 과정을 억제하는 기능을 하고 있음을 발견하였고, 해당 유전자가 녹아웃 된 경우 대조구에 비해 재분화 효율이 현저하게 개선되는 것을 확인함으로써, 본 발명을 완성하였다.
상기 과제를 해결하기 위해, 본 발명은 애기장대 유래 LUX (LUX ARRHYTHMO), IDD7 (INDETERMINATE(ID)-DOMAIN 7), SVP (SHORT VEGETATIVE PHASE), SEP3 (SEPALLATA 3), CALS1 (CALLOSE SYNTHASE 1), WRI1 (WRINKLED 1), KCS2 (3-KETOACYL-COA SYNTHASE 2), KCS20, CRY2 (CRYPTOCHROME 2) 또는 COP1 (CONSTITUTIVE PHOTOMORPHOGENIC 1) 단백질을 코딩하는 유전자의 발현을 식물세포에서 조절하는 단계를 포함하는, 식물체의 재분화(regeneration) 효율을 조절하는 방법을 제공한다.
또한, 본 발명은 애기장대 유래 LUX, IDD7, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질을 코딩하는 유전자를 포함하는 재조합 벡터로 식물세포를 형질전환하는 단계를 포함하는, 재분화 효율이 조절된 형질전환 식물체의 제조방법을 제공한다.
또한, 본 발명은 상기 제조방법에 의해 제조된 재분화 효율이 조절된 형질전환 식물체 및 이의 형질전환된 종자를 제공한다.
또한, 본 발명은 애기장대 유래 LUX, IDD7/11, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질을 코딩하는 유전자를 유효성분으로 포함하는, 식물체의 재분화 효율 조절용 조성물을 제공한다.
또한, 본 발명은 애기장대 유래 LUX, IDD7/11, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질을 코딩하는 유전자의 발현이 저해되어, 야생형에 비해 재분화 효율이 증가된 돌연변이 식물체를 제공한다.
본 발명에 따른 애기장대 유래 LUX, IDD7/11, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 유전자는 식물체의 재분화 효율을 효과적으로 개선할 수 있으므로, 작물 형질전환 효율 증진에 기여할 수 있을 것이며, 육종연한을 단축시키고 육종 과정의 효율화를 가능하게 할 수 있을 것으로 기대된다.
도 1은 야생형(Col-0 또는 Ler)과 녹아웃 돌연변이체(lux, idd7-3, idd7-3xidd11-2, svp-32, sep3-2, cals1, tcp4-ko, wri1-3, kcs2-ko, kcs20-ko, cry2/fha-1, 및 cop1-4)의 하배축 절편체 유래 캘러스를 신초 유도배지에 동일기간 배양하였을 때 형성된 신초의 개수를 보여준다.
본 발명의 목적을 달성하기 위하여, 본 발명은 애기장대 유래 LUX (LUX ARRHYTHMO), IDD7 (INDETERMINATE(ID)-DOMAIN 7), SVP (SHORT VEGETATIVE PHASE), SEP3 (SEPALLATA 3), CALS1 (CALLOSE SYNTHASE 1), WRI1 (WRINKLED 1), KCS2 (3-KETOACYL-COA SYNTHASE 2), KCS20, CRY2 (CRYPTOCHROME 2) 또는 COP1 (CONSTITUTIVE PHOTOMORPHOGENIC 1) 단백질을 코딩하는 유전자의 발현을 식물세포에서 조절하는 단계를 포함하는, 식물체의 재분화(regeneration) 효율을 조절하는 방법을 제공한다.
본 발명의 상기 애기장대 유래 LUX, IDD7, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 및 COP1 단백질은 각각 순서대로 서열번호 12 내지 21로 표시되는 아미노산 서열을 갖는 단백질 및 상기 단백질의 기능적 동등물을 포함한다. "기능적 동등물"이란 아미노산의 부가, 치환 또는 결실의 결과, 상기 서열번호 12 내지 21로 표시되는 아미노산 서열과 적어도 70% 이상, 바람직하게는 80% 이상, 더욱 바람직하게는 90% 이상, 더 더욱 바람직하게는 95% 이상의 서열 상동성을 갖는 것으로, 서열번호 12 내지 21로 표시되는 단백질과 실질적으로 동질의 생리활성을 나타내는 단백질을 말한다. "실질적으로 동질의 생리활성"이란 식물체의 재분화 효율을 조절하는 활성을 의미한다.
또한, 본 발명은 상기 애기장대 유래 LUX, IDD7, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 및 COP1 단백질을 코딩하는 유전자를 제공한다. 상기 유전자는 애기장대 유래 LUX, IDD7, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 및 COP1 단백질을 암호화하는 게놈 DNA와 cDNA를 모두 포함한다. 본 발명의 애기장대 유래 LUX, IDD7, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 및 COP1 단백질을 코딩하는 유전자는 각각 순서대로 서열번호 1 내지 10으로 표시되는 염기서열을 포함할 수 있다. 또한, 상기 염기서열의 상동체가 본 발명의 범위 내에 포함된다. 구체적으로, 상기 애기장대 유래 LUX, IDD7, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 및 COP1 단백질을 코딩하는 유전자는 서열번호 1 내지 10의 염기 서열과 각각 70% 이상, 더욱 바람직하게는 80% 이상, 더 더욱 바람직하게는 90% 이상, 가장 바람직하게는 95% 이상의 서열 상동성을 가지는 염기 서열을 포함할 수 있다. 폴리뉴클레오티드에 대한 "서열 상동성의 %"는 두 개의 최적으로 배열된 서열과 비교 영역을 비교함으로써 확인되며, 비교 영역에서의 폴리뉴클레오티드 서열의 일부는 두 서열의 최적 배열에 대한 참고 서열(추가 또는 삭제를 포함하지 않음)에 비해 추가 또는 삭제(즉, 갭)를 포함할 수 있다.
본 발명의 일 구현 예에 따른 식물체의 재분화 효율 조절 방법에 있어서, 상기 애기장대 유래 LUX, IDD7, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질을 코딩하는 유전자의 발현 조절은 바람직하게는 LUX, IDD7, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질 코딩 유전자의 발현이 저해된 것으로, 식물세포에서 LUX, IDD7, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질 코딩 유전자의 발현 저해는 야생형에 비해 식물체의 재분화 효율을 증가시켰다.
또한, 본 발명의 일 구현 예에 있어서, 상기 IDD7 단백질을 코딩하는 유전자의 발현 저해는 서열번호 22의 아미노산 서열로 이루어진 애기장대 유래 IDD11 단백질을 코딩하는 유전자의 발현 저해를 추가로 포함할 수 있고, IDD7 및 IDD11의 이중 녹아웃(발현 저해)은 IDD7 유전자 단일 놋아웃과 비교하여 재분화 효율이 증가된다.
상기 애기장대 유래 IDD11 단백질을 코딩하는 유전자는 서열번호 11의 염기서열로 이루어진 것일 수 있다.
본 발명의 일 구현 예에 있어서, 상기 LUX, IDD7/11, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질을 코딩하는 유전자의 발현 억제는 상기 각각의 단백질 코딩 유전자에 VIGS (Virus-induced gene silencing) 벡터, RNAi (RNA interference) 또는 안티센스 RNA, T-DNA 삽입, 내생 트랜스포존(transposon), 방사선 조사 또는 유전자 교정 시스템을 이용하여 LUX, IDD7/11, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질 코딩 유전자의 발현을 저해(억제)하는 것일 수 있으나, 이에 제한되지 않으며, 유전자의 발현을 저해하는 당업계의 통상의 방법이면 모두 가능할 수 있다.
또한, 본 발명에 있어서, 상기 증가된 재분화 효율은 지상부 재분화일 수 있고, 구체적으로는 하배축(hypocotyl) 유래 캘러스로부터 신초 형성을 증가시키는 것일 수 있으나, 이에 제한되는 것은 아니다.
본 발명은 또한, 애기장대 유래 LUX, IDD7, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질을 코딩하는 유전자를 포함하는 재조합 벡터로 식물세포를 형질전환하는 단계를 포함하는, 재분화 효율이 조절된 형질전환 식물체의 제조방법을 제공한다.
본 발명의 일 구현 예에 따른 형질전환 식물체의 제조방법은 상기 애기장대 유래 LUX, IDD7, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질을 코딩하는 유전자의 발현을 식물세포에서 저해시켜 야생형에 비해 식물체의 재분화 효율을 증가시키는 것을 특징으로 하나, 이에 제한되지 않는다.
또한, 본 발명의 일 구현 예에 있어서, 상기 IDD7 단백질을 코딩하는 유전자의 발현 저해는 IDD11 단백질을 코딩하는 유전자의 발현 저해를 추가로 포함할 수 있으나, 이에 제한되지 않는다.
본 발명에 따른 형질전환 식물체의 제조방법에 있어서, 상기 LUX, IDD7/11, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 및 COP1 단백질, 및 상기 단백질을 코딩하는 유전자의 범위는 전술한 것과 같다.
본 명세서에서 용어 "재조합"은 세포가 이종의 핵산을 복제하거나, 상기 핵산을 발현하거나 또는 펩티드, 이종의 펩티드 또는 이종의 핵산에 의해 암호된 단백질을 발현하는 세포를 지칭하는 것이다. 재조합 세포는 상기 세포의 천연 형태에서는 발견되지 않는 유전자 또는 유전자 절편을, 센스 또는 안티센스 형태 중 하나로 발현할 수 있다. 또한 재조합 세포는 천연 상태의 세포에서 발견되는 유전자를 발현할 수 있으며, 그러나 상기 유전자는 변형된 것으로서 인위적인 수단에 의해 세포내 재도입된 것이다.
본 명세서에서 용어 "벡터"는 세포 내로 전달하는 DNA 단편(들), 핵산 분자를 지칭할 때 사용된다. 벡터는 DNA를 복제시키고, 숙주세포에서 독립적으로 재생산될 수 있다. 용어 "전달체"는 흔히 "벡터"와 호환하여 사용된다. 용어 "발현 벡터"는 목적한 코딩 서열과, 특정 숙주 생물에서 작동가능하게 연결된 코딩 서열을 발현하는데 필수적인 적정 핵산 서열을 포함하는 재조합 DNA 분자를 의미한다. 진핵세포에서 이용 가능한 프로모터, 인핸서, 종결신호 및 폴리아데닐레이션 신호는 공지되어 있다.
본 발명의 애기장대 유래 LUX, IDD7/11, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질을 코딩하는 유전자 서열 및 적당한 전사/번역 조절 신호를 포함하는 발현 벡터는 당업자에 주지된 방법에 의해 구축될 수 있다. 상기 방법은 시험관내 재조합 DNA 기술, DNA 합성 기술 및 생체 내 재조합 기술 등을 포함한다. 상기 DNA 서열은 mRNA 합성을 이끌기 위해 발현 벡터 내의 적당한 프로모터에 효과적으로 연결될 수 있다. 또한 발현 벡터는 번역 개시 부위로서 리보좀 결합 부위 및 전사 터미네이터를 포함할 수 있다.
본 발명의 재조합 벡터의 바람직한 예는 VIGS 벡터 또는 RNAi 벡터이다. VIGS는 바이러스 벡터에 식물유전자를 도입한 후 식물체를 감염시키면, 그 도입된 유전자의 내인성 식물유전자가 발현이 억제되는 현상을 말한다. 이는 PTGS (Post-transcriptional gene silencing)의 일종으로서, 전사-후 (post-transcriptional), RNA 턴오버 (RNA turnover) 및 뉴클레오티드 서열 특이적 (nucleotide sequence-specific) 이라는 특징들을 가진다. 상기 VIGS 벡터는 외래 유전자를 도입한 식물체 내에서 일시적으로 발현시킬 수 있는 일시적 (transient) 발현 벡터 및 외래 유전자가 도입된 식물체에서 영구적으로 발현시킬 수 있는 식물 발현 벡터로 사용할 수 있다.
식물 발현 벡터의 바람직한 예는 아그로박테리움 투머파시엔스 (Agrobacterium tumefaciens)와 같은 적당한 숙주에 존재할 때 그 자체의 일부, 소위 T-영역을 식물 세포로 전이시킬 수 있는 Ti-플라스미드 벡터이다. 다른 유형의 Ti-플라스미드 벡터 (EP 0 116 718 B1호 참조)는 현재 식물 세포, 또는 잡종 DNA를 식물의 게놈 내에 적당하게 삽입시키는 새로운 식물이 생산될 수 있는 원형질체로 잡종 DNA 서열을 전이시키는데 이용되고 있다. Ti-플라스미드 벡터의 특히 바람직한 형태는 EP 0 120 516 B1호 및 미국 특허 제4,940,838호에 청구된 바와 같은 소위 바이너리 (binary) 벡터이다. 본 발명에 따른 DNA를 식물 숙주에 도입시키는데 이용될 수 있는 다른 적합한 벡터는 이중 가닥 식물 바이러스 (예를 들면, CaMV) 및 단일 가닥 바이러스, 게미니 바이러스 등으로부터 유래될 수 있는 것과 같은 바이러스 벡터, 예를 들면 비완전성 식물 바이러스 벡터로부터 선택될 수 있다. 그러한 벡터의 사용은 특히 식물 숙주를 적당하게 형질전환하는 것이 어려울 때 유리할 수 있다.
발현 벡터는 바람직하게는 하나 이상의 선택성 마커를 포함할 것이다. 상기 마커는 통상적으로 화학적인 방법으로 선택될 수 있는 특성을 갖는 핵산 서열로, 형질전환된 세포를 비형질전환 세포로부터 구별할 수 있는 모든 유전자가 이에 해당된다. 그 예로는 글리포세이트(glyphosate), 포스피노트리신(phosphinothricin) 및 글루포시네이트(glufosinate)와 같은 제초제 저항성 유전자, 카나마이신(kanamycin), G418, 블레오마이신(bleomycin), 하이그로마이신(hygromycin), 클로람페니콜(chloramphenicol)과 같은 항생제 내성 유전자가 있으나, 이에 한정되는 것은 아니다.
본 발명의 식물 발현 벡터에서, 프로모터는 바람직하게는 T7 프로모터, SP6 프로모터, CaMV 35S 프로모터, 액틴 프로모터, 유비퀴틴 프로모터, pEMU 프로모터, MAS 프로모터 또는 히스톤 프로모터일 수 있으나, 이에 제한되지 않는다. 용어 "프로모터"란 구조 유전자로부터의 DNA 업스트림의 영역을 의미하며 전사를 개시하기 위하여 RNA 폴리머라아제가 결합하는 DNA 분자를 말한다. "식물 프로모터"는 식물 세포에서 전사를 개시할 수 있는 프로모터이다. "구성적 (constitutive) 프로모터"는 대부분의 환경 조건 및 발달 상태 또는 세포 분화하에서 활성이 있는 프로모터이다. 형질전환체의 선택이 각종 단계에서 각종 조직에 의해서 이루어질 수 있기 때문에 구성적 프로모터가 본 발명에서 바람직할 수 있다. 따라서, 구성적 프로모터는 선택 가능성을 제한하지 않는다.
본 발명의 재조합 벡터에서, 통상의 터미네이터를 사용할 수 있으며, 그 예로는 노팔린 신타아제(Nopaline synthase), 벼 α-아밀라아제 RAmy1 A 터미네이터, 파세올린(Phaseolin) 터미네이터, 아그로박테리움 투메파시엔스 (A. tumefaciens)의 옥토파인 신타아제(Octopine synthase) 유전자의 터미네이터, 대장균의 rrnB1/B2 터미네이터 등이 있으나, 이에 한정되는 것은 아니다. 터미네이터의 필요성에 관하여, 그러한 영역이 식물 세포에서의 전사의 확실성 및 효율을 증가시키는 것으로 일반적으로 알려져 있다. 그러므로, 터미네이터의 사용은 본 발명의 내용에서 매우 바람직하다.
본 발명의 벡터를 숙주세포 내로 운반하는 방법은 원형질체에 대한 칼슘/폴리에틸렌 글리콜 방법(Krens, F.A. et al., 1982, Nature 296, 72-74; Negrutiu I. et al., June 1987, Plant Mol. Biol. 8, 363-373), 원형질체의 전기천공법(Shillito R.D. et al., 1985 Bio/Technol. 3, 1099-1102), 식물 요소로의 현미주사법(Crossway A. et al., 1986, Mol. Gen. Genet. 202, 179-185), 각종 식물 요소의(DNA 또는 RNA-코팅된) 입자 충격법(Klein T.M. et al., 1987, Nature 327, 70), 식물의 침윤 또는 성숙 화분 또는 소포자의 형질전환에 의한 아그로박테리움 투머파시엔스 매개된 유전자 전이에서(비완전성) 바이러스에 의한 감염(EP 0 301 316호) 등으로부터 적당하게 선택될 수 있다.
또한, 본 발명의 제조방법은 상기 형질전환된 식물세포로부터 형질전환 전식물로 재분화하는 단계를 포함한다. 형질전환 식물세포로부터 형질전환 전식물을 재분화하는 방법은 당업계에 공지된 임의의 방법을 이용할 수 있다.
식물의 형질전환에 이용되는 "식물세포"는 어떤 식물세포도 된다. 식물세포는 배양 세포, 배양 조직, 배양기관 또는 전체 식물이다. "식물 조직"은 분화된 또는 미분화된 식물의 조직, 예를 들면 이에 한정되진 않으나, 뿌리, 줄기, 잎, 꽃가루, 종자, 암 조직 및 배양에 이용되는 다양한 형태의 세포들, 즉 단일 세포, 원형질체(protoplast), 싹 및 캘러스 조직을 포함한다. 식물 조직은 인 플란타(in planta)이거나 기관 배양, 조직배양 또는 세포 배양 상태일 수 있다.
본 발명은 또한, 상기 제조방법에 의해 제조된 재분화 효율이 조절된 형질전환 식물체 및 이의 형질전환된 종자를 제공한다. 바람직하게는, 상기 형질전환 식물체 및 이의 형질전환된 종자는 야생형에 비해 재분화 효율이 증가된 형질전환 식물체 및 이의 형질전환된 종자이다.
상기 식물체는 애기장대, 감자, 가지, 담배, 고추, 토마토, 우엉, 쑥갓, 상추, 도라지, 시금치, 근대, 고구마, 샐러리, 당근, 미나리, 파슬리, 배추, 양배추, 갓무, 수박, 참외, 오이, 호박, 박, 딸기, 대두, 녹두, 강낭콩, 완두 등의 쌍자엽 식물 또는 벼, 보리, 밀, 호밀, 옥수수, 사탕수수, 귀리, 양파 등의 단자엽 식물일 수 있으며, 바람직하게는 쌍자엽 식물이며, 더욱 바람직하게는 애기장대 식물체일 수 있으나, 이에 제한되지 않는다.
본 발명은 또한, 애기장대 유래 LUX, IDD7/11, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질을 코딩하는 유전자를 유효성분으로 포함하는, 식물체의 재분화 효율 조절용 조성물을 제공한다. 본 발명에 따른 조성물은 유효성분으로 식물체의 재분화 효율을 조절할 수 있는 애기장대 유래 LUX, IDD7/11, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질 코딩 유전자를 포함하며, 상기 LUX, IDD7/11, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질 코딩 유전자의 발현이 저해되면 식물체의 재분화 효율을 증가시킬 수 있다.
본 발명에 따른 조성물에 있어서, 상기 IDD7 단백질 코딩 유전자의 발현 저해는 IDD11 단백질 코딩 유전자의 발현 저해를 추가로 포함할 수 있고, IDD7 및 IDD11의 이중 녹아웃(발현 저해)은 IDD7 유전자 단일 놋아웃과 비교하여 재분화 효율이 증가되는 것이 특징이다.
본 발명의 조성물에 있어서, 상기 애기장대 유래 LUX, IDD7/11, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 및 COP1 단백질, 및 상기 단백질을 코딩하는 유전자의 범위는 전술한 것과 같다.
본 발명은 또한, 애기장대 유래 LUX, IDD7/11, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질을 코딩하는 유전자의 발현이 저해되어, 야생형에 비해 재분화 효율이 증가된 돌연변이 식물체를 제공한다. 상기 유전자의 발현 저해는 LUX, IDD7/11, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질을 코딩하는 유전자에 T-DNA 또는 내생 트랜스포존(transposon)을 삽입하여 기능상실 돌연변이체를 유도하거나, LUX, IDD7/11 SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질을 코딩하는 유전자를 포함하는 VIGS (Virus-induced gene silencing) 벡터 또는 RNAi (RNA interference) 벡터를 식물세포에 형질전환하여 LUX, IDD7/11, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 돌연변이체를 유도하여 이루어질 수 있으나, 이에 제한되지 않는다.
본 발명에 따른 돌연변이 식물체에 있어서, 상기 IDD7 단백질 코딩 유전자의 발현이 저해된 돌연변이 식물체는 IDD11 단백질 코딩 유전자의 발현이 저해된 것을 추가로 포함할 수 있고, IDD7 및 IDD11의 이중 녹아웃(발현 저해)은 IDD7 유전자 단일 놋아웃과 비교하여 재분화 효율이 증가된 것이 특징이다.
본 발명의 돌연변이 식물체에 있어서, 상기 애기장대 유래 LUX, IDD7/11, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 및 COP1 단백질, 및 상기 단백질을 코딩하는 유전자의 범위는 전술한 것과 같다.
이하, 본 발명을 실시예에 의해 상세히 설명한다. 단, 하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예에 한정되는 것은 아니다.
재료 및 방법
1. 식물 재료 및 성장 조건
본 발명에서는 Columbia-0 생태형 (Col-0) 또는 Landsberg 생태형 (Ler) 애기장대를 야생형으로 사용하였다. 애기장대 종자의 표면을 멸균한 후, 1/2 MS (Murashige and Skoog)를 포함하는 0.7% 아가 플레이트에 심었다. 식물은 22-23℃에서 백색 형광등 (120 μmol photons·m-2·s-1)을 사용하여 장일조건 (16 시간 광/8 시간 암주기)으로 재배되었다. 본 발명에서 사용된 LUX, IDD7/IDD11, SVP, SEP3, CALS1, TCP4, WRI1, KCS2, KCS20, CRY2 및 COP1 녹아웃 돌연변이체[lux (Salk-022315), idd7-3, idd7-3xidd11-2, svp-32, sep3-2, cals1, tcp4-ko, wri1-3, kcs2-ko, kcs20-ko, cry2/fha-1(CS108), 및 cop1-4]의 출처는 하기 표 1과 같다.
| 돌연변이체 | 출처 |
| lux | ABRC (https://abrc.osu.edu/) 구입, Salk_022315 |
| idd7-3 | 세종대학교 김현욱 교수님 (Salk_129306) |
| idd7-3xidd11-2 | 세종대학교 김현욱 교수님 (idd11-2, Salk_058044) |
| svp-32 | ABRC 구입, Salk_072930C |
| sep3-2 | 성균관대학교 정재훈 교수님 |
| cals1 | ABRC 구입, Salk_142792C |
| tcp4-ko | Spain / Center for Research in Agricultural Genomics의 Dr. Paloma Mas |
| wri1-3 | 세종대학교 김현욱 교수님 (Salk_085693) |
| kcs2-ko | 서강대학교 서미정 교수님 (Salk_033206) |
| kcs20-ko | 서강대학교 서미정 교수님 (Salk_019727) |
| cry2/fha-1 | ABRC 구입, Stock no. CS108 |
| cop1-4 | 서울대학교 백남천 교수님 (돌연변이체 정보: McNellis, T.W. et al., 1994 Plant Cell 6: 487-500) |
2. 지상부 재분화
지상부 재분화(shoot regeneration)를 위해, 7일령 식물체의 하배축 절편체를 캘러스 유도배지 [callus-inducing medium (CIM); 0.5 ㎍/㎖ 2,4-디클로로페녹시아세트산 (2,4-D) 및 0.05 ㎍/㎖ 키네틴이 보충된 MS 배지]에서 22℃의 조건으로 7일간 암배양하였다. 그 후, 상기 CIM에서 7일간 전배양한 캘러스를 신초 유도배지 [shoot-inducing medium (SIM); 0.9 μmol/L 3-인돌아세트산, 2.5 μmol/L 2-이소펜테닐아데닌이 보충된 MS 배지]에서 25℃, 연속광 조건 하에서 18일 동안 배양하였다.
실시예 1. 야생형과 돌연변이체의 지상부 재분화능 분석
식물체의 재분화는 캘러스 유도와 de novo 지상부 기관형성 (organogenesis)을 포함하는 두 단계 과정을 포함한다. 먼저 조직 절편체는 옥신이 풍부한 캘러스 유도배지에 배양하여 분화된 체세포로부터 캘러스 형성을 자극시킨다. 다능성 (pluripotent)의 캘러스는 그 후 옥신 대비 시토키닌 (cytokinin)의 높은 비율을 포함하는 신초 유도배지로 옮겨 de novo 신초 형성을 유도하게 된다. 신초 유도배지에서 배양함에 따라 시토키닌 신호전달의 양성 조절자인 type B-ARR (ARABIDOPSIS RESPONSE REGULATOR)이 신초 줄기 세포 형성의 마스터 조절자인 WUS (WUSCHEL)를 활성화시킨다. 새로운 증거들은 많은 유전적 요인이 시험관내 (in vitro) 식물 재생에 관여한다는 것을 보여주었다. 식물체의 재분화능은 주로 유전적 배경에 의해 결정되기 때문에, 체외식물 재생의 유전적 기여도를 확실히 파악할 필요가 있다.
이에 본 발명자는 다양한 유전자 돌연변이체를 사용하여 신초 재생 능력을 스크리닝하였다. 돌연변이 식물체의 하배축 절편체 유래 캘러스로부터 재생되는 잎을 계산하여 지상부 재분화능을 살펴본 결과, LUX, IDD7, IDD7xIDD11, SVP, SEP3, CALS1, TCP4, WRI1, KCS2, KCS20, CRY2 (=FHA) 및 COP1 유전자의 돌연변이체 유래 캘러스가 야생형 유래 캘러스에 비해 신초 형성 수가 현저히 많은 것을 확인할 수 있었다(도 1). 이의 결과를 통해, 본 발명의 LUX, IDD7/11, SVP, SEP3, CALS1, TCP4, WRI1, KCS2, KCS20, CRY2 및 COP1 유전자는 발현 저해를 통해 재분화 효율을 증진시킬 수 있으므로, 형질전환 또는 유전체 교정 작물의 재분화 효율 증진에 유용하게 활용될 수 있을 것으로 사료되었다.
<110> Seoul National University R&DB Foundation
<120> Gene from Arabidopsis thaliana for regulating regeneration
efficiency of plant and uses thereof
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ccaccgaatt tggcgatggc gtttagtatc acaccggaga gaagccgtac gattcaggat 180
gtgaatcgtg catcggagac gacgctctcg tcgctacgtg gtggatcttc aggtccaaat 240
acctcgtcgt cgaataataa cgtggaggag gaagatcgag ttggatctag cagtcctgga 300
tcggattcga agaagcaaaa gacatcaaac ggtgatggag atgacggtgg cggtgtggat 360
ccggattcgg cgatggcggc ggaagaagga gattcaggaa ctgaagatct atctgggaaa 420
acacttaaac gaccgcgttt agtgtggaca ccgcagctac acaagagatt cgttgacgtt 480
gtagctcact tagggatcaa aaacgctgtt ccgaagacga ttatgcagct gatgaacgtt 540
gaaggattaa ctcgtgagaa cgttgcgtct catcttcaaa agtataggct ttacctcaaa 600
aggatgcagg gattgactaa tgaaggtccc tctgcttcgg ataagctctt ctcttcaaca 660
cctgttcctc cacagagctt ccaagatatc ggtggcggtg gcggtagcag cggtaatgtt 720
ggagtgccga ttccgggggc gtatggaacg caacagatga tgcagatgcc agtttatgca 780
catcatatgg gtatgcaagg atatcatcat caaaatcata atcatgatcc ttatcatcag 840
aatcatcgtc atcatcatgg agctggtgga aatggttctt ggtgtctaca aaatttctta 900
tcttttgcgg agttttccac cgcaaagtcc caaattgcat ctgttcagga gaggatcata 960
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atgtcgaatc taacatcagc ttcaggagat caagcaagtg tgtcttcagg aaacagaact 120
gaaactagtg gctctaacat taatcagcat catcaagaac agtgttttgt tccacaatcg 180
tctcttaaga ggaagagaaa tcaaccagga aatccagacc cagaagcaga agtgatggcc 240
ttatcaccaa agacactaat ggcgacaaac agattcatat gtgaggtctg caacaaaggt 300
tttcaaagag accaaaactt gcaacttcac aagagaggac ataatctacc gtggaagctt 360
aaacaaagat caaacaaaga tgtggttagg aagaaagttt atgtttgtcc agagccaggc 420
tgtgtccatc accatccatc gagagctctt ggtgacttaa ccggaatcaa gaagcatttc 480
ttcagaaaac atggcgagaa gaaatggaaa tgtgaaaagt gttccaagaa atatgcggtg 540
caatcagatt ggaaggctca tgctaagact tgtggcacta aagaatacaa atgcgactgt 600
ggaactctct tttctaggag ggatagtttc ataactcata gagcattttg cgatgcatta 660
gcagaagaga gtgcaagagc catgccaaac cctattatga tccaagcttc aaattctcct 720
catcatcatc atcatcaaac acaacaaaac atcggtttct cttcgtcatc ccaaaacatc 780
atcagcaaca gcaatctcca tggtcctatg aagcaagaag aatcacagca tcactatcaa 840
aatatccctc cttggcttat ctcatcaaac cctaacccta atggcaacaa tggtaatttg 900
ttccctcctg ttgcttcttc tgtgaatacc gggaggtcca gctttcccca cccatcgccg 960
gctatgtcag ccacagcttt gctccagaaa gcagctcaaa tgggttctac gaagtcaaca 1020
actccagagg aagaagagag gtcttcgagg tcaagctata ataacttgat cacgactaca 1080
atggctgcta tgatgacttc accaccagaa cctggcttcg gatttcaaga ctattacatg 1140
atgaatcatc aacatcacgg tggaggagaa gctttcaacg gtggttttgt ccccggagaa 1200
gagaagaacg acgtcgtgga cgacggtgga ggagagacga gagatttctt ggggctaagg 1260
tcgttaatgt cgcataacga gattctaagt ttcgcgaata atcttggcaa ctgtctcaac 1320
acttctgcta cagagcaaca acaacaacaa catagccacc aagattaa 1368
<210> 3
<211> 723
<212> DNA
<213> Arabidopsis thaliana
<400> 3
atggcgagag aaaagattca gatcaggaag atcgacaacg caacggcgag acaagtgacg 60
ttttcgaaac gaagaagagg gcttttcaag aaagctgaag aactctccgt tctctgcgac 120
gccgatgtcg ctctcatcat cttctcttcc accggaaaac tgttcgagtt ctgtagctcc 180
agcatgaagg aagtcctaga gaggcataac ttgcagtcaa agaacttgga gaagcttgat 240
cagccatctc ttgagttaca gctggttgag aacagtgatc acgcccgaat gagtaaagaa 300
attgcggaca agagccaccg actaaggcaa atgagaggag aggaacttca aggacttgac 360
attgaagagc ttcagcagct agagaaggcc cttgaaactg gtttgacgcg tgtgattgaa 420
acaaagagtg acaagattat gagtgagatc agcgaacttc agaaaaaggg aatgcaattg 480
atggatgaga acaagcggtt gaggcagcaa ggaacgcaac taacggaaga gaacgagcga 540
cttggcatgc aaatatgtaa caatgtgcat gcacacggtg gtgctgaatc ggagaacgct 600
gctgtgtacg aggaaggaca gtcgtcggag tctattacta acgccggaaa ctctaccgga 660
gcgcctgttg actccgagag ctccgacact tcccttaggc tcggcttacc gtatggtggt 720
tag 723
<210> 4
<211> 755
<212> DNA
<213> Arabidopsis thaliana
<400> 4
atgggaagag ggagagtaga attgaagagg atagagaaca agatcaatag gcaagtgacg 60
tttgcaaaga gaaggaatgg tcttttgaag aaagcatacg agctttcagt tctatgtgat 120
gcagaagttg ctctcatcat cttctcaaat agaggaaagc tgtacgagtt ttgcagtagt 180
tcgagcatgc ttcggacact ggagaggtac caaaagtgta actatggagc accagaaccc 240
aatgtgcctt caagagaggc cttagcagtt gaacttagta gccagcagga gtatctcaag 300
cttaaggagc gttatgacgc cttacagaga acccaaagga atctgttggg agaagatctt 360
ggacctctaa gtacaaagga gcttgagtca cttgagagac agcttgattc ttccttgaag 420
cagatcagag ctctcaggac acagtttatg cttgaccagc tcaacgatct tcagagtaag 480
gaacgcatgc tgactgagac aataaaactc taagactaag gttagctgat gggtatcaga 540
tgccactcca gctgaaccct aaccaagaag aggttgatca ctacggtcgt catcatcatc 600
aacaacaaca acactcccaa gctttcttcc agcctttgga atgtgaaccc attcttcaga 660
tcgggtatca ggggcaacaa gatggaatgg gagcaggacc aagtgtgaat aattacatgt 720
tgggttggtt accttatgac accaactcta tttga 755
<210> 5
<211> 5853
<212> DNA
<213> Arabidopsis thaliana
<400> 5
atggctcaaa gaagggaacc tgatccgcct ccaccgcaga ggcggattct gcggactcag 60
actgttggta gtctcggaga ggctatgctt gatagtgaag ttgttccctc ttctttagtt 120
gagattgctc cgattctacg tgtggctaat gaagttgaag ctagtaatcc acgagttgct 180
tacctatgtc gtttctacgc atttgagaag gcgcataggc ttgatccaac ttcaagtggt 240
cgtggtgttc gccagtttaa gaccgcgctt cttcaacgat tagaaaggga gaatgaaaca 300
actttggcag gaaggcaaaa gagtgatgca cgtgaaatgc agagtttcta tcaacactac 360
tacaagaaat acatccaagc tttgctgaat gctgcggata aagctgaccg tgctcaactt 420
acaaaggcct atcaaactgc tgccgttctc tttgaagtct tgaaggctgt taatcaaaca 480
gaagatgtgg aagtggctga tgagattttg gaaacgcaca ataaggtgga agagaagacc 540
cagatttacg tgccttacaa tatacttccc cttgaccctg atagtcagaa tcaggcaata 600
atgagattgc ctgagataca agctgctgta gccgctctta gaaatacaag gggattacca 660
tggacagcag gtcacaagaa gaaactcgat gaagacattc tagactggct tcagtctatg 720
tttggttttc agaaagataa tgttttgaat caaagagagc acttgatctt gttacttgct 780
aatgtccaca tccgacaatt tccaaaacct gaccaacaac caaagttgga tgatcgtgcc 840
ttgaccatag tgatgaagaa acttttcaga aactacaaaa agtggtgcaa atacttgggt 900
cggaaaagca gcctttggtt gcctactatt cagcaagagg tacagcagcg taaactgctc 960
tacatgggac tttatctatt aatatgggga gaagctgcaa acctgagatt catgccagaa 1020
tgcctctgtt atatatatca ccatatggct ttcgagcttt atggtatgtt ggccggtagt 1080
gttagtccca tgacagggga gcatgtaaag ccagcgtatg gtggtgaaga tgaagctttc 1140
ctgcagaaag ttgtgactcc tatataccag acaatatcaa aggaagcaaa gagaagtaga 1200
ggaggaaagt caaagcattc tgtgtggaga aactatgatg acttgaatga atatttttgg 1260
tcaattcgat gttttcgctt agggtggcca atgcgggctg atgctgattt cttttgtcag 1320
acagctgaag aacttcgact tgagagaagt gagataaaat caaacagtgg agatcggtgg 1380
atgggcaaag tcaattttgt ggagatacgc tcgttctggc atatatttag aagttttgat 1440
agattgtgga gcttttatat cctgtgtttg caggcaatga ttgttattgc atggaatgga 1500
tccggagagt taagcgccat ttttcagggt gatgtcttcc taaaggtttt gagcgtcttc 1560
attacagcag ctatactaaa gcttgcacaa gctgtccttg acatagccct cagctggaag 1620
gcaagacata gtatgtcact ctatgtgaaa ctcagatacg tcatgaaggt aggtgcggct 1680
gcagtatggg ttgtagtcat ggcagttact tatgcgtaca gctggaaaaa cgcctccggt 1740
ttttctcaga ccataaagaa ttggtttggg ggacacagcc ataattcacc ttctcttttc 1800
atagtggcaa ttcttatcta cttatccccg aacatgcttt ctgcgttgct gtttttgttt 1860
ccattcattc ggcgctatct tgaaaggtct gactataaga taatgatgct gatgatgtgg 1920
tggtctcagc ctcgtctgta tataggaagg ggtatgcatg agagcgcttt gtcacttttc 1980
aagtacacaa tgttttggat cgttctgctg atttcaaagc tggcattcag ttattacgct 2040
gagattaagc cccttgttgg tccaaccaaa gatataatgc gaatccatat atccgtctac 2100
agctggcatg agtttttccc tcacgccaag aataatttag gagtcgtcat cgcgctctgg 2160
tcaccagtta ttctcgttta tttcatggac actcaaatat ggtatgctat agtttcaact 2220
ctggtcggag gtttaaatgg agctttccgc cgtcttggag agatccgaac acttggaatg 2280
ttgaggtcga gatttcagtc cataccaggg gcctttaatg attgcttggt accacaagat 2340
aacagcgatg atacaaagaa aaagagattt agggcgacct tctctcgcaa gtttgatcag 2400
ctcccatcta gtaaagacaa ggaagcagcg cggtttgctc agatgtggaa caaaataatc 2460
agtagtttcc gagaagagga cttgattagt gatagggaaa tggaactatt gcttgtgcca 2520
tattggtctg accctgattt ggatcttatc cggtggccac cttttctatt ggctagtaag 2580
atcccaatag ctttggatat ggctaaagac agcaatggaa aagaccgtga actgaaaaag 2640
agattagctg ttgacagcta catgacttgt gcagttagag agtgctatgc ctcttttaag 2700
aatctcatca attatttggt ggtcggagaa cgagaaggac aggtcataaa cgacattttc 2760
tccaagattg atgaacatat tgagaaggaa acgctgataa cagaattgaa cctgagtgct 2820
cttcctgatc tgtatggcca gttcgttcgg ctaatcgaat atttgctaga aaacagggag 2880
gaggacaagg atcaaattgt aattgtcttg ctgaacatgt tagaattggt gactagagac 2940
atcatggaag aagaagtccc aagcttacta gagactgccc acaatggatc ttatgtcaag 3000
tatgacgtca tgacccctct tcatcaacag cgcaaatatt tcagtcaact tcgattccca 3060
gtttattctc aaacggaagc ttggaaagaa aaaattaaga ggcttcatct tttgcttact 3120
gtcaaggagt cagctatgga tgttccatcc aacttggaag ctcgaaggcg tcttactttc 3180
ttttcaaatt cactattcat ggacatgcct cctgcaccca agatccgcaa catgctttcc 3240
ttctcggtcc ttacaccata cttttcagag gatgttcttt tctccatatt tggcttagaa 3300
cagcaaaatg aagatggagt ttctatactc ttttacttgc agaagatctt cccagatgaa 3360
tggacaaact ttctggaacg agttaaatgt ggcaacgaag aagaactaag agcaagagag 3420
gatttggaag aggagcttcg tctttgggca tcttacagag gccaaactct gactaaaact 3480
gtgcgaggaa tgatgtatta ccgaaaggct ttggaacttc aggccttcct tgatatggca 3540
aaggatgaag aattattaaa aggctacaag gctctcgagt taaccagtga agaagcatca 3600
aagagcggag ggtcactttg ggcgcagtgt caagcacttg cagatatgaa gttcaccttt 3660
gtggtttctt gtcaacaata cagtattcat aagagatctg gcgaccaacg tgctaaagac 3720
atattgagac ttatgacgac gtatccgtct attcgtgtcg cctatatcga tgaggtagag 3780
caaacacata aagaaagtta taaggggaca gaggagaaaa tttattactc agcgcttgta 3840
aaagctgctc cacagaccaa accaatggat tcttctgaat ctgttcaaac actagatcag 3900
ctcatttatc ggattaagct tccgggacca gctatactag gagagggaaa gccggaaaat 3960
cagaaccatg ctatcatatt tacccgtgga gaagggttgc agacgataga catgaatcag 4020
gacaactaca tggaggaagc tttcaaaatg aggaacttgc tgcaagagtt tcttgaaaag 4080
catggaggcg taagatgtcc tacgattctt ggtcttagag agcatatttt cactggaagt 4140
gtgtcttctc ttgcatggtt tatgtcaaat caagagaaca gttttgtaac gattgggcaa 4200
agagtgctag ctagtccctt gaaagtacga ttccattacg gacatccaga tatttttgat 4260
cgtctgtttc accttaccag aggtggtatc tgcaaagctt ctaaagtcat caatcttagt 4320
gaagacatat ttgcaggttt taattctact ctgcgtgaag gaaatgtgac tcatcatgaa 4380
tatatacaag ttggtaaagg gagagatgtg ggcctcaacc agatctcaat gtttgaggca 4440
aaaatcgcca atggaaatgg cgagcaaact ttgagtcgcg acctttatag gctaggacac 4500
cgatttgatt tcttccggat gctgtcttgt tatttcacca caattgggtt ctacttcagt 4560
accatgttaa ccgtgcttac ggtgtatgtg tttctttatg gacggttata tctagtactt 4620
agtgggcttg aagaaggatt gagtagccaa cgagctttcc gtaacaataa gcctcttgag 4680
gcagctcttg cttctcaatc atttgtgcag attggttttt taatggcttt gcctatgatg 4740
atggagattg gacttgagag ggggttccac aatgcattga ttgagtttgt gctgatgcag 4800
ttacaacttg cttccgtctt tttcacgttt cagctcggga caaaaacaca ttactacgga 4860
aggacattgt ttcatggagg cgcagagtat agaggaactg gtcgagggtt tgtggtcttc 4920
cacgcgaaat ttgctgagaa ctataggttc tattcccgta gtcactttgt gaagggcatt 4980
gaacttatga ttcttctact ggtatatcag atctttggtc aatcatatag gggtgtggtt 5040
acttatattc tgatcactgt ttcgatatgg ttcatggtgg tgacatggct ctttgctcct 5100
ttccttttca atccgtctgg ttttgagtgg caaaagattg ttgatgactg gacagactgg 5160
aacaaatgga tttataaccg aggaggaatt ggtgtccctc ctgagaagag ttgggaatct 5220
tggtgggaga aagagttaga acatcttaga cattcagggg ttcgtggtat cacccttgag 5280
atctttttgg ccttgcgctt cttcatcttt caatatggac ttgtgtatca tctcagtacc 5340
ttcaaaggaa aaaatcagag cttttgggtt tatggagcgt catggtttgt gatcttattc 5400
attctactga ttgtgaaggg tttgggcgtt ggaaggcgaa gattcagcac gaatttccag 5460
cttctattca gaatcataaa gggtcttgtg ttcctcacat ttgtggcaat tctcataact 5520
ttcctcgcac ttcctctgat aacaatcaaa gacttattca tctgcatgct tgccttcatg 5580
ccaactggat ggggaatgct tctcattgca caagcgtgta agcctctgat ccaacagcta 5640
gggatatggt catcagttag gacgctagct cgaggctacg agatcgtgat gggcttgctt 5700
ctgttcacac cagttgcgtt tttggcctgg tttccttttg tgtctgagtt ccagacaaga 5760
atgctcttta accaagcctt tagtagaggt ctacaaatct ctcgtattct tggtggtcaa 5820
agaaaagacc ggtcttcaaa aaacaaggag tga 5853
<210> 6
<211> 1317
<212> DNA
<213> Arabidopsis thaliana
<400> 6
atgaagaagc gcttaaccac ttccacttgt tcttcttctc catcttcctc tgtttcttct 60
tctactacta cttcctctcc tattcagtcg gaggctccaa ggcctaaacg agccaaaagg 120
gctaagaaat cttctccttc tggtgataaa tctcataacc cgacaagccc tgcttctacc 180
cgacgcagct ctatctacag aggagtcact agacatagat ggactgggag attcgaggct 240
catctttggg acaaaagctc ttggaattcg attcagaaca agaaaggcaa acaagtttat 300
ctgggagcat atgacagtga agaagcagca gcacatacgt acgatctggc tgctctcaag 360
tactggggac ccgacaccat cttgaatttt ccggcagaga cgtacacaaa ggaattggaa 420
gaaatgcaga gagtgacaaa ggaagaatat ttggcttctc tccgccgcca gagcagtggt 480
ttctccagag gcgtctctaa atatcgcggc gtcgctaggc atcaccacaa cggaagatgg 540
gaggctcgga tcggaagagt gtttgggaac aagtacttgt acctcggcac ctataatacg 600
caggaggaag ctgctgcagc atatgacatg gctgcgattg agtatcgagg cgcaaacgcg 660
gttactaatt tcgacattag taattacatt gaccggttaa agaagaaagg tgttttcccg 720
ttccctgtga accaagctaa ccatcaagag ggtattcttg ttgaagccaa acaagaagtt 780
gaaacgagag aagcgaagga agagcctaga gaagaagtga aacaacagta cgtggaagaa 840
ccaccgcaag aagaagaaga gaaggaagaa gagaaagcag agcaacaaga agcagagatt 900
gtaggatatt cagaagaagc agcagtggtc aattgctgca tagactcttc aaccataatg 960
gaaatggatc gttgtgggga caacaatgag ctggcttgga acttctgtat gatggataca 1020
gggttttctc cgtttttgac tgatcagaat ctcgcgaatg agaatcccat agagtatccg 1080
gagctattca atgagttagc atttgaggac aacatcgact tcatgttcga tgatgggaag 1140
cacgagtgct tgaacttgga aaatctggat tgttgcgtgg tgggaagaga gagcccaccc 1200
tcttcttctt caccattgtc ttgcttatct actgactctg cttcatcaac aacaacaaca 1260
acaacctcgg tttcttgtaa ctatttgttt cagggcttgt tcgttggttc tgaataa 1317
<210> 7
<211> 1587
<212> DNA
<213> Arabidopsis thaliana
<400> 7
atgaatgaga atcacattca aagtgatcac atgaacaaca ccattcatgt aacaaacaaa 60
aaactcccaa acttcctctt atccgtacgt ctcaaatatg ttaaacttgg ttaccattac 120
ctaatctcga atgccgtcta cattctcata ctccccgtcg gcctcctcgc cgcaacctcc 180
tcctccttta gtctcaccga tctcacactc ttatacaacc accttctcaa attccacttc 240
ctctcttcca ctctcttcgc cgctcttctt atctttctaa ccaccctcta cttcacaact 300
cgtcctcgca gaatctttct cctcgatttc gcttgttaca aacccgactc ttctctaata 360
tgcacacgag aaacgttcat ggaccggtct cagcgtgtcg gtatcttcac agaagacaac 420
ctcgctttcc aacaaaagat cctcgaacgt tccggtcttg gacaaaagac ttacttcccg 480
gaggctctct tacgtgtccc gcctaatcct tgtatgtctg aagcaaggaa agaagccgag 540
acggttatgt ttggagctat agacgcagtg cttgagaaaa ccggagtgaa tcctaaggat 600
attgggatac ttgtcgtgaa ttgtagcttg tttaatccga cgccttcgtt atcggctatg 660
atcgtgaata aatataagct cagaggaaac gttttgagct ataaccttgg tggaatgggt 720
tgtagtgctg gtcttatctc cattgatctc gctaaacagc ttcttcaggt tcaaccaaat 780
tcatatgcac tagtagtgag cacagagaac ataaccctaa actggtattt aggcaacgac 840
cgatcaatgc ttctctcaaa ctgcatcttc cgaatgggtg gcgccgccgt tctcctctct 900
aaccgctcct ccgaccgctg ccgctccaaa taccaactca tccacaccgt ccgtacacac 960
aaaggatctg acgacaacgc ctttaactgc gtttaccaac gcgaagacaa cgacgacaac 1020
aaacaaatcg gtgtctcact ttccaaaaac ctaatggcta tagctggaga agctctcaag 1080
acgaacataa ccactctggg accgctagtt ctaccaatgt ctgaacaact tctctttttc 1140
gcaacactag tggcgcgaaa agtctttaat gtgaagaaaa taaaacctta tattcctgat 1200
ttcaagctag ctttcgagca cttctgcatc cacgcgggag gcagagcagt gctcgatgag 1260
attgagaaga atttggattt atctgagtgg catatggaac cttcgaggat gactttgaac 1320
cggtttggaa atacttcgag tagctcgctt tggtatgagc ttgcgtatag tgaggctaaa 1380
ggaaggatta agagaggaga taggacttgg cagattgctt ttggatccgg ttttaagtgt 1440
aatagtgcgg tttggagagc tttgagaact atcgatccgt ccaaggagaa gaagaagaag 1500
actaatcctt ggattgatga gattcatgag tttcccgtgc cggttccaag gacatctccg 1560
gtgacatctt cctcggaatc tcgatag 1587
<210> 8
<211> 1590
<212> DNA
<213> Arabidopsis thaliana
<400> 8
atgagccata accaaaacca acctcaccgg ccagtcccgg tccatgtcac aaacgccgag 60
ccaaacccta acccaaacaa cctcccaaac tttctattat ccgtccgtct caaatacgta 120
aaactcggtt accactacct aatctccaac gctctctaca tcctcctcct tcctctcctc 180
gccgcaacaa tcgctaacct ctcttctttc accatcaacg acctctctct cctctacaac 240
acactccgtt tccatttcct ctccgccaca ctcgccaccg cactcttaat ctctctctcc 300
accgcttact tcaccacccg tccccgccgt gtcttcctcc tcgacttctc gtgttacaaa 360
ccagaccctt cactgatctg cactcgtgaa acattcatgg acagatctca acgtgtaggc 420
atcttcacag aagacaactt agctttccaa caaaagatcc tcgaaagatc cggtctaggt 480
cagaaaactt acttccctga agctcttctt cgtgttcctc ctaatccttg tatggaagaa 540
gcgagaaaag aggcagaaac agttatgttc ggagctattg acgcggttct tgagaagacc 600
ggtgtgaaac ctaaagatat tggaatcctt gtggtgaatt gtagcttgtt taatccaaca 660
ccgtcacttt ctgctatgat tgtgaataag tataagctta gaggcaacat tttgagctat 720
aatcttggtg gaatgggatg tagtgctggc cttatctcca ttgatctcgc taaacagatg 780
cttcaggtgc aaccaaactc atacgcacta gtggtgagca cagagaacat aaccctaaac 840
tggtacttag gcaacgaccg atcaatgctt ctatctaact gtatcttccg tatgggcgga 900
gccgcggtac ttctctcgaa ccgctcctct gatcgcagcc gttcaaaata tcagctcatc 960
cataccgtcc gtacccacaa aggagctgat gacaacgcat ttggctgtgt ttaccaacga 1020
gaagacaaca acgcagaaga aaccggcaaa atcggagtgt ccctctctaa aaacctaatg 1080
gcgatagctg gagaagctct caagacaaac atcacaactc tcggaccact agttctacca 1140
atgtccgaac aacttctctt tttcgcgact cttgtggccc gaaaagtctt caaagtcaag 1200
aaaataaagc cttacattcc tgatttcaag ctagctttcg agcatttctg tatccacgcg 1260
ggaggtagag ccgtgctcga cgagattgag aagaacttgg atttatccga atggcacatg 1320
gagccatcga ggatgacgtt gaaccggttt ggtaacactt cgagtagctc actttggtat 1380
gagcttgcgt atagtgaagc taaaggaagg attaagagag gagataggac ttggcagatt 1440
gcttttggat cgggttttaa gtgtaatagt gcagtttgga aagctttgag aacgattgat 1500
ccaatggacg agaagactaa tccatggatt gatgagattg atgactttcc tgttcaagtt 1560
cctaggatca ctcccattac atcgtcgtag 1590
<210> 9
<211> 1839
<212> DNA
<213> Arabidopsis thaliana
<400> 9
atgaagatgg acaaaaagac tatagtttgg tttagaagag acctaaggat tgaggataat 60
cctgcattag cagcagctgc tcacgaagga tctgtttttc ctgtcttcat ttggtgtcct 120
gaagaagaag gacagtttta tcctggaaga gcttcaagat ggtggatgaa acaatcactt 180
gctcacttat ctcaatcctt gaaggctctt ggatctgacc tcactttaat caaaacccac 240
aacacgattt cagcgatctt ggattgtatc cgcgttaccg gtgctacaaa agtcgtcttt 300
aaccacctct atgatcctgt ttcgttagtt cgggaccata ccgtaaagga gaagctggtg 360
gaacgtggga tctctgtgca aagctacaat ggagatctat tgtatgaacc gtgggagata 420
tactgcgaaa agggcaaacc ttttacgagt ttcaattctt actggaagaa atgcttagat 480
atgtcgattg aatccgttat gcttcctcct ccttggcggt tgatgccaat aactgcagcg 540
gctgaagcga tttgggcgtg ttcgattgaa gaactagggc tggagaatga ggccgagaaa 600
ccgagcaatg cgttgttaac tagagcttgg tctccaggat ggagcaatgc tgataagtta 660
ctaaatgagt tcatcgagaa gcagttgata gattatgcaa agaacagcaa gaaagttgtt 720
gggaattcta cttcactact ttctccgtat ctccatttcg gggaaataag cgtcagacac 780
gttttccagt gtgcccggat gaaacaaatt atatgggcaa gagataagaa cagtgaagga 840
gaagaaagtg cagatctttt tcttagggga atcggtttaa gagagtattc tcggtatata 900
tgtttcaact tcccgtttac tcacgagcaa tcgttgttga gtcatcttcg gtttttccct 960
tgggatgctg atgttgataa gttcaaggcc tggagacaag gcaggaccgg ttatccgttg 1020
gtggatgccg gaatgagaga gctttgggct accggatgga tgcataacag aataagagtg 1080
attgtttcaa gctttgctgt gaagtttctt ctccttccat ggaaatgggg aatgaagtat 1140
ttctgggata cacttttgga tgctgatttg gaatgtgaca tccttggctg gcagtatatc 1200
tctgggagta tccccgatgg ccacgagctt gatcgcttgg acaatcccgc gttacaaggc 1260
gccaaatatg acccagaagg tgagtacata aggcaatggc ttcccgagct tgcgagattg 1320
ccaactgaat ggatccatca tccatgggac gctcctttaa ccgtactcaa agcttctggt 1380
gtggaactcg gaacaaacta tgcgaaaccc attgtagaca tcgacacagc tcgtgagcta 1440
ctagctaaag ctatttcaag aacccgtgaa gcacagatca tgatcggagc agcacctgat 1500
gagattgtag cagatagctt cgaggcctta ggggctaata ccattaaaga acctggtctt 1560
tgcccatctg tgtcttctaa tgaccaacaa gtaccttcgg ctgttcgtta caacgggtca 1620
aagagagtga aacctgagga agaagaagag agagacatga agaaatctag gggattcgat 1680
gaaagggagt tgttttcgac tgctgaatct tcttcttctt cgagtgtgtt tttcgtttcg 1740
cagtcttgct cgttggcatc agaagggaag aatctggaag gtattcaaga ttcatctgat 1800
cagattacta caagtttggg aaaaaatggt tgcaaatga 1839
<210> 10
<211> 2028
<212> DNA
<213> Arabidopsis thaliana
<400> 10
atggaagaga tttcgacgga tccggttgtt ccagcggtga aacctgaccc gagaacatct 60
tcagttggtg aaggtgctaa tcgtcatgaa aatgacgacg gaggaagcgg cggttctgag 120
attggagcac cggatctgga taaagacttg ctttgtccga tttgtatgca gattattaaa 180
gatgctttcc tcacggcttg tggtcatagt ttctgctata tgtgtatcat cacacatctt 240
aggaacaaga gtgattgtcc ctgttgtagc caacacctca ccaataatca gctttaccct 300
aatttcttgc tcgataagct attgaagaaa acttcagctc ggcatgtgtc aaaaactgca 360
tcgcccttgg atcagtttcg ggaagcacta caaaggggtt gtgatgtgtc aattaaggag 420
gttgataatc ttctgacact tcttgcggaa aggaagagaa aaatggaaca ggaagaagct 480
gagaggaaca tgcagatact tttggacttt ttgcattgtc taaggaagca aaaagttgat 540
gaactaaatg aggtgcaaac tgatctccag tatattaaag aagatataaa tgccgttgag 600
agacatagaa tagatttata ccgagctagg gacagatatt ctgtaaagtt gcggatgctc 660
ggagatgatc caagcacaag aaatgcatgg ccacatgaga agaaccagat tggtttcaac 720
tccaattctc tcagcataag aggaggaaat tttgtaggca attatcaaaa caaaaaggta 780
gaggggaagg cacaaggaag ctctcatggg ctaccaaaga aggatgcgct gagtgggtca 840
gattcgcaaa gtttgaatca gtcaactgtc tcaatggcta gaaagaaacg gattcatgct 900
cagttcaatg atttacaaga atgttacctc caaaagcggc gtcagttggc agaccaacca 960
aatagtaaac aagaaaatga taagagtgta gtacggaggg aaggctatag caacggcctt 1020
gcagattttc aatctgtgtt gactaccttc actcgctaca gtcgtctaag agttatagca 1080
gaaatccggc atggggatat atttcattca gccaacattg tatcaagcat agagtttgat 1140
cgtgatgatg agctgtttgc cactgctggt gtttctagat gtataaaggt ttttgacttc 1200
tcttcggttg taaatgaacc agcagatatg cagtgtccga ttgtggagat gtcaactcgg 1260
tctaaactta gttgcttgag ttggaataag catgaaaaaa atcacatagc aagcagtgat 1320
tatgaaggaa tagtaacagt gtgggatgta actactaggc agagtcttat ggagtatgaa 1380
gagcacgaaa aacgtgcctg gagtgttgac ttttcacgaa cagaaccatc aatgcttgta 1440
tctggtagtg acgactgcaa ggttaaagtt tggtgcacga ggcaggaagc aagtgtgatt 1500
aatattgata tgaaagcaaa catatgttgt gtcaagtaca atcctggctc aagcaactac 1560
attgcggtcg gatcagctga tcatcacatc cattattacg atctaagaaa cataagccaa 1620
ccacttcatg tcttcagtgg acacaagaaa gcagtttcct atgttaaatt tttgtccaac 1680
aacgagctcg cttctgcgtc cacagatagc acactacgct tatgggatgt caaagacaac 1740
ttgccagttc gaacattcag aggacatact aacgagaaga actttgtggg tctcacagtg 1800
aacagcgagt atctcgcctg tggaagcgag acaaacgaag tatatgtata tcacaaggaa 1860
atcacgagac ccgtgacatc gcacagattt ggatcgccag acatggacga tgcagaggaa 1920
gaggcaggtt cctactttat tagtgcggtt tgctggaaga gtgatagtcc cacgatgttg 1980
actgcgaata gtcaaggaac catcaaagtt ctggtactcg ctgcgtga 2028
<210> 11
<211> 1545
<212> DNA
<213> Arabidopsis thaliana
<400> 11
atgttacttc accagcatca gcaaccacaa caagacgaga atatgtcgaa tctaacatca 60
gcttccgggg atcaagcaag tgtctcttca ggaaatataa ctgaagcaag tggttctaat 120
tacttccctc atcatcagca acagcaagaa caacaacaac aacaaataca aaaactttca 180
tgctcttgga ctgattcatt attccaatta tttgatacag tcaccttctt ggaaattctg 240
tacccagaat cagaagtgat agctctatca ccaaaaacac taatggcaac aaacagattc 300
gtgtgtgaaa tctgcaacaa aggttttcaa agagaccaga atctacagct tcatagaaga 360
ggtcacaatc tgccatggaa gctgaaacaa agatccaaca aagaagtgat aaggaagaaa 420
gtgtatgtat gtccagaagc aagctgtgtc caccatgatc catctagagc tctcggtgac 480
ttaaccggaa tcaagaagca tttctgtaga aaacatggtg agaagaagtg gaaatgtgat 540
aagtgttcaa agaaatatgc tgttcaatca gattgcaagg ctcattctaa gacttgtggc 600
accaaagaat acagatgtga ctgtggcact ctcttttcta ggagggatag tttcataact 660
catagagcat tttgtgaagc attggctgaa gagactgcaa gagaagtagt aataccacaa 720
aaccagaata ataatcaacc aaaccctctt ttgattcatc aatctgcttc tcatcctcat 780
caccatcatc aaacacaacc aaccataaac gtctcttctt cttcatcttc ctcccacaac 840
cacaacatca tcaatagcct tcacttcgac accaacaatg gtaatactaa taatagtaac 900
aacagcaaca accatctcca tacatttccc atgaagaaag agcaacaaag caatgaccat 960
atcatgaatt accaccatag tatcatccct ccttggcttg ctcctcagcc tcacgcactc 1020
acatcttcaa accctaaccc tagtaatggt ggaggaggag gaggaagttt gttctctctt 1080
gcttctccgg ctatgtcagc caccgcattg ctccagaaag cagcccaaat gggctctaca 1140
aagacacctc ctttaccacc aaccacggcc tacgagaggt caactcataa taacaacctc 1200
acaacgacaa tggcggcgat gatgacgtca ccatctggat tcatcagctc caacaacaac 1260
aatcatgttt tgtttcaaga ctacaacgca tccgggtttg ataatcacgg cagagaagaa 1320
gcattcgacg atacatttgg cgggttcttg agaacaaacg aagttacagc cgccgcagga 1380
tcagagaaga gtactaaaag cggtggagga gaaggtttga cgagagattt cttggggcta 1440
agaccgttaa tgtctcataa cgagattcta agttttgctg gtcttggaag ttgcataaat 1500
agctctgctt ctgatcagct tcatccaaag ccttggcagg gttag 1545
<210> 12
<211> 324
<212> PRT
<213> Arabidopsis thaliana
<400> 12
Met Gly Glu Glu Val Gln Met Ser Asp Tyr Asp Val Ser Gly Asp Gly
1 5 10 15
Asp Arg Val Ser Glu Trp Glu Met Gly Leu Pro Ser Asp Glu Asp Leu
20 25 30
Ala Ser Leu Ser Tyr Ser Leu Ile Pro Pro Asn Leu Ala Met Ala Phe
35 40 45
Ser Ile Thr Pro Glu Arg Ser Arg Thr Ile Gln Asp Val Asn Arg Ala
50 55 60
Ser Glu Thr Thr Leu Ser Ser Leu Arg Gly Gly Ser Ser Gly Pro Asn
65 70 75 80
Thr Ser Ser Ser Asn Asn Asn Val Glu Glu Glu Asp Arg Val Gly Ser
85 90 95
Ser Ser Pro Gly Ser Asp Ser Lys Lys Gln Lys Thr Ser Asn Gly Asp
100 105 110
Gly Asp Asp Gly Gly Gly Val Asp Pro Asp Ser Ala Met Ala Ala Glu
115 120 125
Glu Gly Asp Ser Gly Thr Glu Asp Leu Ser Gly Lys Thr Leu Lys Arg
130 135 140
Pro Arg Leu Val Trp Thr Pro Gln Leu His Lys Arg Phe Val Asp Val
145 150 155 160
Val Ala His Leu Gly Ile Lys Asn Ala Val Pro Lys Thr Ile Met Gln
165 170 175
Leu Met Asn Val Glu Gly Leu Thr Arg Glu Asn Val Ala Ser His Leu
180 185 190
Gln Lys Tyr Arg Leu Tyr Leu Lys Arg Met Gln Gly Leu Thr Asn Glu
195 200 205
Gly Pro Ser Ala Ser Asp Lys Leu Phe Ser Ser Thr Pro Val Pro Pro
210 215 220
Gln Ser Phe Gln Asp Ile Gly Gly Gly Gly Gly Ser Ser Gly Asn Val
225 230 235 240
Gly Val Pro Ile Pro Gly Ala Tyr Gly Thr Gln Gln Met Met Gln Met
245 250 255
Pro Val Tyr Ala His His Met Gly Met Gln Gly Tyr His His Gln Asn
260 265 270
His Asn His Asp Pro Tyr His Gln Asn His Arg His His His Gly Ala
275 280 285
Gly Gly Asn Gly Ser Trp Cys Leu Gln Asn Phe Leu Ser Phe Ala Glu
290 295 300
Phe Ser Thr Ala Lys Ser Gln Ile Ala Ser Val Gln Glu Arg Ile Ile
305 310 315 320
Gln Ser Ile Met
<210> 13
<211> 455
<212> PRT
<213> Arabidopsis thaliana
<400> 13
Met Met Met Asn Arg Asp Ile Leu Phe His Gln Gln Gln Gln Gln Gln
1 5 10 15
Met Glu Glu Asn Met Ser Asn Leu Thr Ser Ala Ser Gly Asp Gln Ala
20 25 30
Ser Val Ser Ser Gly Asn Arg Thr Glu Thr Ser Gly Ser Asn Ile Asn
35 40 45
Gln His His Gln Glu Gln Cys Phe Val Pro Gln Ser Ser Leu Lys Arg
50 55 60
Lys Arg Asn Gln Pro Gly Asn Pro Asp Pro Glu Ala Glu Val Met Ala
65 70 75 80
Leu Ser Pro Lys Thr Leu Met Ala Thr Asn Arg Phe Ile Cys Glu Val
85 90 95
Cys Asn Lys Gly Phe Gln Arg Asp Gln Asn Leu Gln Leu His Lys Arg
100 105 110
Gly His Asn Leu Pro Trp Lys Leu Lys Gln Arg Ser Asn Lys Asp Val
115 120 125
Val Arg Lys Lys Val Tyr Val Cys Pro Glu Pro Gly Cys Val His His
130 135 140
His Pro Ser Arg Ala Leu Gly Asp Leu Thr Gly Ile Lys Lys His Phe
145 150 155 160
Phe Arg Lys His Gly Glu Lys Lys Trp Lys Cys Glu Lys Cys Ser Lys
165 170 175
Lys Tyr Ala Val Gln Ser Asp Trp Lys Ala His Ala Lys Thr Cys Gly
180 185 190
Thr Lys Glu Tyr Lys Cys Asp Cys Gly Thr Leu Phe Ser Arg Arg Asp
195 200 205
Ser Phe Ile Thr His Arg Ala Phe Cys Asp Ala Leu Ala Glu Glu Ser
210 215 220
Ala Arg Ala Met Pro Asn Pro Ile Met Ile Gln Ala Ser Asn Ser Pro
225 230 235 240
His His His His His Gln Thr Gln Gln Asn Ile Gly Phe Ser Ser Ser
245 250 255
Ser Gln Asn Ile Ile Ser Asn Ser Asn Leu His Gly Pro Met Lys Gln
260 265 270
Glu Glu Ser Gln His His Tyr Gln Asn Ile Pro Pro Trp Leu Ile Ser
275 280 285
Ser Asn Pro Asn Pro Asn Gly Asn Asn Gly Asn Leu Phe Pro Pro Val
290 295 300
Ala Ser Ser Val Asn Thr Gly Arg Ser Ser Phe Pro His Pro Ser Pro
305 310 315 320
Ala Met Ser Ala Thr Ala Leu Leu Gln Lys Ala Ala Gln Met Gly Ser
325 330 335
Thr Lys Ser Thr Thr Pro Glu Glu Glu Glu Arg Ser Ser Arg Ser Ser
340 345 350
Tyr Asn Asn Leu Ile Thr Thr Thr Met Ala Ala Met Met Thr Ser Pro
355 360 365
Pro Glu Pro Gly Phe Gly Phe Gln Asp Tyr Tyr Met Met Asn His Gln
370 375 380
His His Gly Gly Gly Glu Ala Phe Asn Gly Gly Phe Val Pro Gly Glu
385 390 395 400
Glu Lys Asn Asp Val Val Asp Asp Gly Gly Gly Glu Thr Arg Asp Phe
405 410 415
Leu Gly Leu Arg Ser Leu Met Ser His Asn Glu Ile Leu Ser Phe Ala
420 425 430
Asn Asn Leu Gly Asn Cys Leu Asn Thr Ser Ala Thr Glu Gln Gln Gln
435 440 445
Gln Gln His Ser His Gln Asp
450 455
<210> 14
<211> 240
<212> PRT
<213> Arabidopsis thaliana
<400> 14
Met Ala Arg Glu Lys Ile Gln Ile Arg Lys Ile Asp Asn Ala Thr Ala
1 5 10 15
Arg Gln Val Thr Phe Ser Lys Arg Arg Arg Gly Leu Phe Lys Lys Ala
20 25 30
Glu Glu Leu Ser Val Leu Cys Asp Ala Asp Val Ala Leu Ile Ile Phe
35 40 45
Ser Ser Thr Gly Lys Leu Phe Glu Phe Cys Ser Ser Ser Met Lys Glu
50 55 60
Val Leu Glu Arg His Asn Leu Gln Ser Lys Asn Leu Glu Lys Leu Asp
65 70 75 80
Gln Pro Ser Leu Glu Leu Gln Leu Val Glu Asn Ser Asp His Ala Arg
85 90 95
Met Ser Lys Glu Ile Ala Asp Lys Ser His Arg Leu Arg Gln Met Arg
100 105 110
Gly Glu Glu Leu Gln Gly Leu Asp Ile Glu Glu Leu Gln Gln Leu Glu
115 120 125
Lys Ala Leu Glu Thr Gly Leu Thr Arg Val Ile Glu Thr Lys Ser Asp
130 135 140
Lys Ile Met Ser Glu Ile Ser Glu Leu Gln Lys Lys Gly Met Gln Leu
145 150 155 160
Met Asp Glu Asn Lys Arg Leu Arg Gln Gln Gly Thr Gln Leu Thr Glu
165 170 175
Glu Asn Glu Arg Leu Gly Met Gln Ile Cys Asn Asn Val His Ala His
180 185 190
Gly Gly Ala Glu Ser Glu Asn Ala Ala Val Tyr Glu Glu Gly Gln Ser
195 200 205
Ser Glu Ser Ile Thr Asn Ala Gly Asn Ser Thr Gly Ala Pro Val Asp
210 215 220
Ser Glu Ser Ser Asp Thr Ser Leu Arg Leu Gly Leu Pro Tyr Gly Gly
225 230 235 240
<210> 15
<211> 251
<212> PRT
<213> Arabidopsis thaliana
<400> 15
Met Gly Arg Gly Arg Val Glu Leu Lys Arg Ile Glu Asn Lys Ile Asn
1 5 10 15
Arg Gln Val Thr Phe Ala Lys Arg Arg Asn Gly Leu Leu Lys Lys Ala
20 25 30
Tyr Glu Leu Ser Val Leu Cys Asp Ala Glu Val Ala Leu Ile Ile Phe
35 40 45
Ser Asn Arg Gly Lys Leu Tyr Glu Phe Cys Ser Ser Ser Ser Met Leu
50 55 60
Arg Thr Leu Glu Arg Tyr Gln Lys Cys Asn Tyr Gly Ala Pro Glu Pro
65 70 75 80
Asn Val Pro Ser Arg Glu Ala Leu Ala Val Glu Leu Ser Ser Gln Gln
85 90 95
Glu Tyr Leu Lys Leu Lys Glu Arg Tyr Asp Ala Leu Gln Arg Thr Gln
100 105 110
Arg Asn Leu Leu Gly Glu Asp Leu Gly Pro Leu Ser Thr Lys Glu Leu
115 120 125
Glu Ser Leu Glu Arg Gln Leu Asp Ser Ser Leu Lys Gln Ile Arg Ala
130 135 140
Leu Arg Thr Gln Phe Met Leu Asp Gln Leu Asn Asp Leu Gln Ser Lys
145 150 155 160
Glu Arg Met Leu Thr Glu Thr Asn Lys Thr Leu Arg Leu Arg Leu Ala
165 170 175
Asp Gly Tyr Gln Met Pro Leu Gln Leu Asn Pro Asn Gln Glu Glu Val
180 185 190
Asp His Tyr Gly Arg His His His Gln Gln Gln Gln His Ser Gln Ala
195 200 205
Phe Phe Gln Pro Leu Glu Cys Glu Pro Ile Leu Gln Ile Gly Tyr Gln
210 215 220
Gly Gln Gln Asp Gly Met Gly Ala Gly Pro Ser Val Asn Asn Tyr Met
225 230 235 240
Leu Gly Trp Leu Pro Tyr Asp Thr Asn Ser Ile
245 250
<210> 16
<211> 1950
<212> PRT
<213> Arabidopsis thaliana
<400> 16
Met Ala Gln Arg Arg Glu Pro Asp Pro Pro Pro Pro Gln Arg Arg Ile
1 5 10 15
Leu Arg Thr Gln Thr Val Gly Ser Leu Gly Glu Ala Met Leu Asp Ser
20 25 30
Glu Val Val Pro Ser Ser Leu Val Glu Ile Ala Pro Ile Leu Arg Val
35 40 45
Ala Asn Glu Val Glu Ala Ser Asn Pro Arg Val Ala Tyr Leu Cys Arg
50 55 60
Phe Tyr Ala Phe Glu Lys Ala His Arg Leu Asp Pro Thr Ser Ser Gly
65 70 75 80
Arg Gly Val Arg Gln Phe Lys Thr Ala Leu Leu Gln Arg Leu Glu Arg
85 90 95
Glu Asn Glu Thr Thr Leu Ala Gly Arg Gln Lys Ser Asp Ala Arg Glu
100 105 110
Met Gln Ser Phe Tyr Gln His Tyr Tyr Lys Lys Tyr Ile Gln Ala Leu
115 120 125
Leu Asn Ala Ala Asp Lys Ala Asp Arg Ala Gln Leu Thr Lys Ala Tyr
130 135 140
Gln Thr Ala Ala Val Leu Phe Glu Val Leu Lys Ala Val Asn Gln Thr
145 150 155 160
Glu Asp Val Glu Val Ala Asp Glu Ile Leu Glu Thr His Asn Lys Val
165 170 175
Glu Glu Lys Thr Gln Ile Tyr Val Pro Tyr Asn Ile Leu Pro Leu Asp
180 185 190
Pro Asp Ser Gln Asn Gln Ala Ile Met Arg Leu Pro Glu Ile Gln Ala
195 200 205
Ala Val Ala Ala Leu Arg Asn Thr Arg Gly Leu Pro Trp Thr Ala Gly
210 215 220
His Lys Lys Lys Leu Asp Glu Asp Ile Leu Asp Trp Leu Gln Ser Met
225 230 235 240
Phe Gly Phe Gln Lys Asp Asn Val Leu Asn Gln Arg Glu His Leu Ile
245 250 255
Leu Leu Leu Ala Asn Val His Ile Arg Gln Phe Pro Lys Pro Asp Gln
260 265 270
Gln Pro Lys Leu Asp Asp Arg Ala Leu Thr Ile Val Met Lys Lys Leu
275 280 285
Phe Arg Asn Tyr Lys Lys Trp Cys Lys Tyr Leu Gly Arg Lys Ser Ser
290 295 300
Leu Trp Leu Pro Thr Ile Gln Gln Glu Val Gln Gln Arg Lys Leu Leu
305 310 315 320
Tyr Met Gly Leu Tyr Leu Leu Ile Trp Gly Glu Ala Ala Asn Leu Arg
325 330 335
Phe Met Pro Glu Cys Leu Cys Tyr Ile Tyr His His Met Ala Phe Glu
340 345 350
Leu Tyr Gly Met Leu Ala Gly Ser Val Ser Pro Met Thr Gly Glu His
355 360 365
Val Lys Pro Ala Tyr Gly Gly Glu Asp Glu Ala Phe Leu Gln Lys Val
370 375 380
Val Thr Pro Ile Tyr Gln Thr Ile Ser Lys Glu Ala Lys Arg Ser Arg
385 390 395 400
Gly Gly Lys Ser Lys His Ser Val Trp Arg Asn Tyr Asp Asp Leu Asn
405 410 415
Glu Tyr Phe Trp Ser Ile Arg Cys Phe Arg Leu Gly Trp Pro Met Arg
420 425 430
Ala Asp Ala Asp Phe Phe Cys Gln Thr Ala Glu Glu Leu Arg Leu Glu
435 440 445
Arg Ser Glu Ile Lys Ser Asn Ser Gly Asp Arg Trp Met Gly Lys Val
450 455 460
Asn Phe Val Glu Ile Arg Ser Phe Trp His Ile Phe Arg Ser Phe Asp
465 470 475 480
Arg Leu Trp Ser Phe Tyr Ile Leu Cys Leu Gln Ala Met Ile Val Ile
485 490 495
Ala Trp Asn Gly Ser Gly Glu Leu Ser Ala Ile Phe Gln Gly Asp Val
500 505 510
Phe Leu Lys Val Leu Ser Val Phe Ile Thr Ala Ala Ile Leu Lys Leu
515 520 525
Ala Gln Ala Val Leu Asp Ile Ala Leu Ser Trp Lys Ala Arg His Ser
530 535 540
Met Ser Leu Tyr Val Lys Leu Arg Tyr Val Met Lys Val Gly Ala Ala
545 550 555 560
Ala Val Trp Val Val Val Met Ala Val Thr Tyr Ala Tyr Ser Trp Lys
565 570 575
Asn Ala Ser Gly Phe Ser Gln Thr Ile Lys Asn Trp Phe Gly Gly His
580 585 590
Ser His Asn Ser Pro Ser Leu Phe Ile Val Ala Ile Leu Ile Tyr Leu
595 600 605
Ser Pro Asn Met Leu Ser Ala Leu Leu Phe Leu Phe Pro Phe Ile Arg
610 615 620
Arg Tyr Leu Glu Arg Ser Asp Tyr Lys Ile Met Met Leu Met Met Trp
625 630 635 640
Trp Ser Gln Pro Arg Leu Tyr Ile Gly Arg Gly Met His Glu Ser Ala
645 650 655
Leu Ser Leu Phe Lys Tyr Thr Met Phe Trp Ile Val Leu Leu Ile Ser
660 665 670
Lys Leu Ala Phe Ser Tyr Tyr Ala Glu Ile Lys Pro Leu Val Gly Pro
675 680 685
Thr Lys Asp Ile Met Arg Ile His Ile Ser Val Tyr Ser Trp His Glu
690 695 700
Phe Phe Pro His Ala Lys Asn Asn Leu Gly Val Val Ile Ala Leu Trp
705 710 715 720
Ser Pro Val Ile Leu Val Tyr Phe Met Asp Thr Gln Ile Trp Tyr Ala
725 730 735
Ile Val Ser Thr Leu Val Gly Gly Leu Asn Gly Ala Phe Arg Arg Leu
740 745 750
Gly Glu Ile Arg Thr Leu Gly Met Leu Arg Ser Arg Phe Gln Ser Ile
755 760 765
Pro Gly Ala Phe Asn Asp Cys Leu Val Pro Gln Asp Asn Ser Asp Asp
770 775 780
Thr Lys Lys Lys Arg Phe Arg Ala Thr Phe Ser Arg Lys Phe Asp Gln
785 790 795 800
Leu Pro Ser Ser Lys Asp Lys Glu Ala Ala Arg Phe Ala Gln Met Trp
805 810 815
Asn Lys Ile Ile Ser Ser Phe Arg Glu Glu Asp Leu Ile Ser Asp Arg
820 825 830
Glu Met Glu Leu Leu Leu Val Pro Tyr Trp Ser Asp Pro Asp Leu Asp
835 840 845
Leu Ile Arg Trp Pro Pro Phe Leu Leu Ala Ser Lys Ile Pro Ile Ala
850 855 860
Leu Asp Met Ala Lys Asp Ser Asn Gly Lys Asp Arg Glu Leu Lys Lys
865 870 875 880
Arg Leu Ala Val Asp Ser Tyr Met Thr Cys Ala Val Arg Glu Cys Tyr
885 890 895
Ala Ser Phe Lys Asn Leu Ile Asn Tyr Leu Val Val Gly Glu Arg Glu
900 905 910
Gly Gln Val Ile Asn Asp Ile Phe Ser Lys Ile Asp Glu His Ile Glu
915 920 925
Lys Glu Thr Leu Ile Thr Glu Leu Asn Leu Ser Ala Leu Pro Asp Leu
930 935 940
Tyr Gly Gln Phe Val Arg Leu Ile Glu Tyr Leu Leu Glu Asn Arg Glu
945 950 955 960
Glu Asp Lys Asp Gln Ile Val Ile Val Leu Leu Asn Met Leu Glu Leu
965 970 975
Val Thr Arg Asp Ile Met Glu Glu Glu Val Pro Ser Leu Leu Glu Thr
980 985 990
Ala His Asn Gly Ser Tyr Val Lys Tyr Asp Val Met Thr Pro Leu His
995 1000 1005
Gln Gln Arg Lys Tyr Phe Ser Gln Leu Arg Phe Pro Val Tyr Ser Gln
1010 1015 1020
Thr Glu Ala Trp Lys Glu Lys Ile Lys Arg Leu His Leu Leu Leu Thr
1025 1030 1035 1040
Val Lys Glu Ser Ala Met Asp Val Pro Ser Asn Leu Glu Ala Arg Arg
1045 1050 1055
Arg Leu Thr Phe Phe Ser Asn Ser Leu Phe Met Asp Met Pro Pro Ala
1060 1065 1070
Pro Lys Ile Arg Asn Met Leu Ser Phe Ser Val Leu Thr Pro Tyr Phe
1075 1080 1085
Ser Glu Asp Val Leu Phe Ser Ile Phe Gly Leu Glu Gln Gln Asn Glu
1090 1095 1100
Asp Gly Val Ser Ile Leu Phe Tyr Leu Gln Lys Ile Phe Pro Asp Glu
1105 1110 1115 1120
Trp Thr Asn Phe Leu Glu Arg Val Lys Cys Gly Asn Glu Glu Glu Leu
1125 1130 1135
Arg Ala Arg Glu Asp Leu Glu Glu Glu Leu Arg Leu Trp Ala Ser Tyr
1140 1145 1150
Arg Gly Gln Thr Leu Thr Lys Thr Val Arg Gly Met Met Tyr Tyr Arg
1155 1160 1165
Lys Ala Leu Glu Leu Gln Ala Phe Leu Asp Met Ala Lys Asp Glu Glu
1170 1175 1180
Leu Leu Lys Gly Tyr Lys Ala Leu Glu Leu Thr Ser Glu Glu Ala Ser
1185 1190 1195 1200
Lys Ser Gly Gly Ser Leu Trp Ala Gln Cys Gln Ala Leu Ala Asp Met
1205 1210 1215
Lys Phe Thr Phe Val Val Ser Cys Gln Gln Tyr Ser Ile His Lys Arg
1220 1225 1230
Ser Gly Asp Gln Arg Ala Lys Asp Ile Leu Arg Leu Met Thr Thr Tyr
1235 1240 1245
Pro Ser Ile Arg Val Ala Tyr Ile Asp Glu Val Glu Gln Thr His Lys
1250 1255 1260
Glu Ser Tyr Lys Gly Thr Glu Glu Lys Ile Tyr Tyr Ser Ala Leu Val
1265 1270 1275 1280
Lys Ala Ala Pro Gln Thr Lys Pro Met Asp Ser Ser Glu Ser Val Gln
1285 1290 1295
Thr Leu Asp Gln Leu Ile Tyr Arg Ile Lys Leu Pro Gly Pro Ala Ile
1300 1305 1310
Leu Gly Glu Gly Lys Pro Glu Asn Gln Asn His Ala Ile Ile Phe Thr
1315 1320 1325
Arg Gly Glu Gly Leu Gln Thr Ile Asp Met Asn Gln Asp Asn Tyr Met
1330 1335 1340
Glu Glu Ala Phe Lys Met Arg Asn Leu Leu Gln Glu Phe Leu Glu Lys
1345 1350 1355 1360
His Gly Gly Val Arg Cys Pro Thr Ile Leu Gly Leu Arg Glu His Ile
1365 1370 1375
Phe Thr Gly Ser Val Ser Ser Leu Ala Trp Phe Met Ser Asn Gln Glu
1380 1385 1390
Asn Ser Phe Val Thr Ile Gly Gln Arg Val Leu Ala Ser Pro Leu Lys
1395 1400 1405
Val Arg Phe His Tyr Gly His Pro Asp Ile Phe Asp Arg Leu Phe His
1410 1415 1420
Leu Thr Arg Gly Gly Ile Cys Lys Ala Ser Lys Val Ile Asn Leu Ser
1425 1430 1435 1440
Glu Asp Ile Phe Ala Gly Phe Asn Ser Thr Leu Arg Glu Gly Asn Val
1445 1450 1455
Thr His His Glu Tyr Ile Gln Val Gly Lys Gly Arg Asp Val Gly Leu
1460 1465 1470
Asn Gln Ile Ser Met Phe Glu Ala Lys Ile Ala Asn Gly Asn Gly Glu
1475 1480 1485
Gln Thr Leu Ser Arg Asp Leu Tyr Arg Leu Gly His Arg Phe Asp Phe
1490 1495 1500
Phe Arg Met Leu Ser Cys Tyr Phe Thr Thr Ile Gly Phe Tyr Phe Ser
1505 1510 1515 1520
Thr Met Leu Thr Val Leu Thr Val Tyr Val Phe Leu Tyr Gly Arg Leu
1525 1530 1535
Tyr Leu Val Leu Ser Gly Leu Glu Glu Gly Leu Ser Ser Gln Arg Ala
1540 1545 1550
Phe Arg Asn Asn Lys Pro Leu Glu Ala Ala Leu Ala Ser Gln Ser Phe
1555 1560 1565
Val Gln Ile Gly Phe Leu Met Ala Leu Pro Met Met Met Glu Ile Gly
1570 1575 1580
Leu Glu Arg Gly Phe His Asn Ala Leu Ile Glu Phe Val Leu Met Gln
1585 1590 1595 1600
Leu Gln Leu Ala Ser Val Phe Phe Thr Phe Gln Leu Gly Thr Lys Thr
1605 1610 1615
His Tyr Tyr Gly Arg Thr Leu Phe His Gly Gly Ala Glu Tyr Arg Gly
1620 1625 1630
Thr Gly Arg Gly Phe Val Val Phe His Ala Lys Phe Ala Glu Asn Tyr
1635 1640 1645
Arg Phe Tyr Ser Arg Ser His Phe Val Lys Gly Ile Glu Leu Met Ile
1650 1655 1660
Leu Leu Leu Val Tyr Gln Ile Phe Gly Gln Ser Tyr Arg Gly Val Val
1665 1670 1675 1680
Thr Tyr Ile Leu Ile Thr Val Ser Ile Trp Phe Met Val Val Thr Trp
1685 1690 1695
Leu Phe Ala Pro Phe Leu Phe Asn Pro Ser Gly Phe Glu Trp Gln Lys
1700 1705 1710
Ile Val Asp Asp Trp Thr Asp Trp Asn Lys Trp Ile Tyr Asn Arg Gly
1715 1720 1725
Gly Ile Gly Val Pro Pro Glu Lys Ser Trp Glu Ser Trp Trp Glu Lys
1730 1735 1740
Glu Leu Glu His Leu Arg His Ser Gly Val Arg Gly Ile Thr Leu Glu
1745 1750 1755 1760
Ile Phe Leu Ala Leu Arg Phe Phe Ile Phe Gln Tyr Gly Leu Val Tyr
1765 1770 1775
His Leu Ser Thr Phe Lys Gly Lys Asn Gln Ser Phe Trp Val Tyr Gly
1780 1785 1790
Ala Ser Trp Phe Val Ile Leu Phe Ile Leu Leu Ile Val Lys Gly Leu
1795 1800 1805
Gly Val Gly Arg Arg Arg Phe Ser Thr Asn Phe Gln Leu Leu Phe Arg
1810 1815 1820
Ile Ile Lys Gly Leu Val Phe Leu Thr Phe Val Ala Ile Leu Ile Thr
1825 1830 1835 1840
Phe Leu Ala Leu Pro Leu Ile Thr Ile Lys Asp Leu Phe Ile Cys Met
1845 1850 1855
Leu Ala Phe Met Pro Thr Gly Trp Gly Met Leu Leu Ile Ala Gln Ala
1860 1865 1870
Cys Lys Pro Leu Ile Gln Gln Leu Gly Ile Trp Ser Ser Val Arg Thr
1875 1880 1885
Leu Ala Arg Gly Tyr Glu Ile Val Met Gly Leu Leu Leu Phe Thr Pro
1890 1895 1900
Val Ala Phe Leu Ala Trp Phe Pro Phe Val Ser Glu Phe Gln Thr Arg
1905 1910 1915 1920
Met Leu Phe Asn Gln Ala Phe Ser Arg Gly Leu Gln Ile Ser Arg Ile
1925 1930 1935
Leu Gly Gly Gln Arg Lys Asp Arg Ser Ser Lys Asn Lys Glu
1940 1945 1950
<210> 17
<211> 438
<212> PRT
<213> Arabidopsis thaliana
<400> 17
Met Lys Lys Arg Leu Thr Thr Ser Thr Cys Ser Ser Ser Pro Ser Ser
1 5 10 15
Ser Val Ser Ser Ser Thr Thr Thr Ser Ser Pro Ile Gln Ser Glu Ala
20 25 30
Pro Arg Pro Lys Arg Ala Lys Arg Ala Lys Lys Ser Ser Pro Ser Gly
35 40 45
Asp Lys Ser His Asn Pro Thr Ser Pro Ala Ser Thr Arg Arg Ser Ser
50 55 60
Ile Tyr Arg Gly Val Thr Arg His Arg Trp Thr Gly Arg Phe Glu Ala
65 70 75 80
His Leu Trp Asp Lys Ser Ser Trp Asn Ser Ile Gln Asn Lys Lys Gly
85 90 95
Lys Gln Val Tyr Leu Gly Ala Tyr Asp Ser Glu Glu Ala Ala Ala His
100 105 110
Thr Tyr Asp Leu Ala Ala Leu Lys Tyr Trp Gly Pro Asp Thr Ile Leu
115 120 125
Asn Phe Pro Ala Glu Thr Tyr Thr Lys Glu Leu Glu Glu Met Gln Arg
130 135 140
Val Thr Lys Glu Glu Tyr Leu Ala Ser Leu Arg Arg Gln Ser Ser Gly
145 150 155 160
Phe Ser Arg Gly Val Ser Lys Tyr Arg Gly Val Ala Arg His His His
165 170 175
Asn Gly Arg Trp Glu Ala Arg Ile Gly Arg Val Phe Gly Asn Lys Tyr
180 185 190
Leu Tyr Leu Gly Thr Tyr Asn Thr Gln Glu Glu Ala Ala Ala Ala Tyr
195 200 205
Asp Met Ala Ala Ile Glu Tyr Arg Gly Ala Asn Ala Val Thr Asn Phe
210 215 220
Asp Ile Ser Asn Tyr Ile Asp Arg Leu Lys Lys Lys Gly Val Phe Pro
225 230 235 240
Phe Pro Val Asn Gln Ala Asn His Gln Glu Gly Ile Leu Val Glu Ala
245 250 255
Lys Gln Glu Val Glu Thr Arg Glu Ala Lys Glu Glu Pro Arg Glu Glu
260 265 270
Val Lys Gln Gln Tyr Val Glu Glu Pro Pro Gln Glu Glu Glu Glu Lys
275 280 285
Glu Glu Glu Lys Ala Glu Gln Gln Glu Ala Glu Ile Val Gly Tyr Ser
290 295 300
Glu Glu Ala Ala Val Val Asn Cys Cys Ile Asp Ser Ser Thr Ile Met
305 310 315 320
Glu Met Asp Arg Cys Gly Asp Asn Asn Glu Leu Ala Trp Asn Phe Cys
325 330 335
Met Met Asp Thr Gly Phe Ser Pro Phe Leu Thr Asp Gln Asn Leu Ala
340 345 350
Asn Glu Asn Pro Ile Glu Tyr Pro Glu Leu Phe Asn Glu Leu Ala Phe
355 360 365
Glu Asp Asn Ile Asp Phe Met Phe Asp Asp Gly Lys His Glu Cys Leu
370 375 380
Asn Leu Glu Asn Leu Asp Cys Cys Val Val Gly Arg Glu Ser Pro Pro
385 390 395 400
Ser Ser Ser Ser Pro Leu Ser Cys Leu Ser Thr Asp Ser Ala Ser Ser
405 410 415
Thr Thr Thr Thr Thr Thr Ser Val Ser Cys Asn Tyr Leu Phe Gln Gly
420 425 430
Leu Phe Val Gly Ser Glu
435
<210> 18
<211> 528
<212> PRT
<213> Arabidopsis thaliana
<400> 18
Met Asn Glu Asn His Ile Gln Ser Asp His Met Asn Asn Thr Ile His
1 5 10 15
Val Thr Asn Lys Lys Leu Pro Asn Phe Leu Leu Ser Val Arg Leu Lys
20 25 30
Tyr Val Lys Leu Gly Tyr His Tyr Leu Ile Ser Asn Ala Val Tyr Ile
35 40 45
Leu Ile Leu Pro Val Gly Leu Leu Ala Ala Thr Ser Ser Ser Phe Ser
50 55 60
Leu Thr Asp Leu Thr Leu Leu Tyr Asn His Leu Leu Lys Phe His Phe
65 70 75 80
Leu Ser Ser Thr Leu Phe Ala Ala Leu Leu Ile Phe Leu Thr Thr Leu
85 90 95
Tyr Phe Thr Thr Arg Pro Arg Arg Ile Phe Leu Leu Asp Phe Ala Cys
100 105 110
Tyr Lys Pro Asp Ser Ser Leu Ile Cys Thr Arg Glu Thr Phe Met Asp
115 120 125
Arg Ser Gln Arg Val Gly Ile Phe Thr Glu Asp Asn Leu Ala Phe Gln
130 135 140
Gln Lys Ile Leu Glu Arg Ser Gly Leu Gly Gln Lys Thr Tyr Phe Pro
145 150 155 160
Glu Ala Leu Leu Arg Val Pro Pro Asn Pro Cys Met Ser Glu Ala Arg
165 170 175
Lys Glu Ala Glu Thr Val Met Phe Gly Ala Ile Asp Ala Val Leu Glu
180 185 190
Lys Thr Gly Val Asn Pro Lys Asp Ile Gly Ile Leu Val Val Asn Cys
195 200 205
Ser Leu Phe Asn Pro Thr Pro Ser Leu Ser Ala Met Ile Val Asn Lys
210 215 220
Tyr Lys Leu Arg Gly Asn Val Leu Ser Tyr Asn Leu Gly Gly Met Gly
225 230 235 240
Cys Ser Ala Gly Leu Ile Ser Ile Asp Leu Ala Lys Gln Leu Leu Gln
245 250 255
Val Gln Pro Asn Ser Tyr Ala Leu Val Val Ser Thr Glu Asn Ile Thr
260 265 270
Leu Asn Trp Tyr Leu Gly Asn Asp Arg Ser Met Leu Leu Ser Asn Cys
275 280 285
Ile Phe Arg Met Gly Gly Ala Ala Val Leu Leu Ser Asn Arg Ser Ser
290 295 300
Asp Arg Cys Arg Ser Lys Tyr Gln Leu Ile His Thr Val Arg Thr His
305 310 315 320
Lys Gly Ser Asp Asp Asn Ala Phe Asn Cys Val Tyr Gln Arg Glu Asp
325 330 335
Asn Asp Asp Asn Lys Gln Ile Gly Val Ser Leu Ser Lys Asn Leu Met
340 345 350
Ala Ile Ala Gly Glu Ala Leu Lys Thr Asn Ile Thr Thr Leu Gly Pro
355 360 365
Leu Val Leu Pro Met Ser Glu Gln Leu Leu Phe Phe Ala Thr Leu Val
370 375 380
Ala Arg Lys Val Phe Asn Val Lys Lys Ile Lys Pro Tyr Ile Pro Asp
385 390 395 400
Phe Lys Leu Ala Phe Glu His Phe Cys Ile His Ala Gly Gly Arg Ala
405 410 415
Val Leu Asp Glu Ile Glu Lys Asn Leu Asp Leu Ser Glu Trp His Met
420 425 430
Glu Pro Ser Arg Met Thr Leu Asn Arg Phe Gly Asn Thr Ser Ser Ser
435 440 445
Ser Leu Trp Tyr Glu Leu Ala Tyr Ser Glu Ala Lys Gly Arg Ile Lys
450 455 460
Arg Gly Asp Arg Thr Trp Gln Ile Ala Phe Gly Ser Gly Phe Lys Cys
465 470 475 480
Asn Ser Ala Val Trp Arg Ala Leu Arg Thr Ile Asp Pro Ser Lys Glu
485 490 495
Lys Lys Lys Lys Thr Asn Pro Trp Ile Asp Glu Ile His Glu Phe Pro
500 505 510
Val Pro Val Pro Arg Thr Ser Pro Val Thr Ser Ser Ser Glu Ser Arg
515 520 525
<210> 19
<211> 529
<212> PRT
<213> Arabidopsis thaliana
<400> 19
Met Ser His Asn Gln Asn Gln Pro His Arg Pro Val Pro Val His Val
1 5 10 15
Thr Asn Ala Glu Pro Asn Pro Asn Pro Asn Asn Leu Pro Asn Phe Leu
20 25 30
Leu Ser Val Arg Leu Lys Tyr Val Lys Leu Gly Tyr His Tyr Leu Ile
35 40 45
Ser Asn Ala Leu Tyr Ile Leu Leu Leu Pro Leu Leu Ala Ala Thr Ile
50 55 60
Ala Asn Leu Ser Ser Phe Thr Ile Asn Asp Leu Ser Leu Leu Tyr Asn
65 70 75 80
Thr Leu Arg Phe His Phe Leu Ser Ala Thr Leu Ala Thr Ala Leu Leu
85 90 95
Ile Ser Leu Ser Thr Ala Tyr Phe Thr Thr Arg Pro Arg Arg Val Phe
100 105 110
Leu Leu Asp Phe Ser Cys Tyr Lys Pro Asp Pro Ser Leu Ile Cys Thr
115 120 125
Arg Glu Thr Phe Met Asp Arg Ser Gln Arg Val Gly Ile Phe Thr Glu
130 135 140
Asp Asn Leu Ala Phe Gln Gln Lys Ile Leu Glu Arg Ser Gly Leu Gly
145 150 155 160
Gln Lys Thr Tyr Phe Pro Glu Ala Leu Leu Arg Val Pro Pro Asn Pro
165 170 175
Cys Met Glu Glu Ala Arg Lys Glu Ala Glu Thr Val Met Phe Gly Ala
180 185 190
Ile Asp Ala Val Leu Glu Lys Thr Gly Val Lys Pro Lys Asp Ile Gly
195 200 205
Ile Leu Val Val Asn Cys Ser Leu Phe Asn Pro Thr Pro Ser Leu Ser
210 215 220
Ala Met Ile Val Asn Lys Tyr Lys Leu Arg Gly Asn Ile Leu Ser Tyr
225 230 235 240
Asn Leu Gly Gly Met Gly Cys Ser Ala Gly Leu Ile Ser Ile Asp Leu
245 250 255
Ala Lys Gln Met Leu Gln Val Gln Pro Asn Ser Tyr Ala Leu Val Val
260 265 270
Ser Thr Glu Asn Ile Thr Leu Asn Trp Tyr Leu Gly Asn Asp Arg Ser
275 280 285
Met Leu Leu Ser Asn Cys Ile Phe Arg Met Gly Gly Ala Ala Val Leu
290 295 300
Leu Ser Asn Arg Ser Ser Asp Arg Ser Arg Ser Lys Tyr Gln Leu Ile
305 310 315 320
His Thr Val Arg Thr His Lys Gly Ala Asp Asp Asn Ala Phe Gly Cys
325 330 335
Val Tyr Gln Arg Glu Asp Asn Asn Ala Glu Glu Thr Gly Lys Ile Gly
340 345 350
Val Ser Leu Ser Lys Asn Leu Met Ala Ile Ala Gly Glu Ala Leu Lys
355 360 365
Thr Asn Ile Thr Thr Leu Gly Pro Leu Val Leu Pro Met Ser Glu Gln
370 375 380
Leu Leu Phe Phe Ala Thr Leu Val Ala Arg Lys Val Phe Lys Val Lys
385 390 395 400
Lys Ile Lys Pro Tyr Ile Pro Asp Phe Lys Leu Ala Phe Glu His Phe
405 410 415
Cys Ile His Ala Gly Gly Arg Ala Val Leu Asp Glu Ile Glu Lys Asn
420 425 430
Leu Asp Leu Ser Glu Trp His Met Glu Pro Ser Arg Met Thr Leu Asn
435 440 445
Arg Phe Gly Asn Thr Ser Ser Ser Ser Leu Trp Tyr Glu Leu Ala Tyr
450 455 460
Ser Glu Ala Lys Gly Arg Ile Lys Arg Gly Asp Arg Thr Trp Gln Ile
465 470 475 480
Ala Phe Gly Ser Gly Phe Lys Cys Asn Ser Ala Val Trp Lys Ala Leu
485 490 495
Arg Thr Ile Asp Pro Met Asp Glu Lys Thr Asn Pro Trp Ile Asp Glu
500 505 510
Ile Asp Asp Phe Pro Val Gln Val Pro Arg Ile Thr Pro Ile Thr Ser
515 520 525
Ser
<210> 20
<211> 612
<212> PRT
<213> Arabidopsis thaliana
<400> 20
Met Lys Met Asp Lys Lys Thr Ile Val Trp Phe Arg Arg Asp Leu Arg
1 5 10 15
Ile Glu Asp Asn Pro Ala Leu Ala Ala Ala Ala His Glu Gly Ser Val
20 25 30
Phe Pro Val Phe Ile Trp Cys Pro Glu Glu Glu Gly Gln Phe Tyr Pro
35 40 45
Gly Arg Ala Ser Arg Trp Trp Met Lys Gln Ser Leu Ala His Leu Ser
50 55 60
Gln Ser Leu Lys Ala Leu Gly Ser Asp Leu Thr Leu Ile Lys Thr His
65 70 75 80
Asn Thr Ile Ser Ala Ile Leu Asp Cys Ile Arg Val Thr Gly Ala Thr
85 90 95
Lys Val Val Phe Asn His Leu Tyr Asp Pro Val Ser Leu Val Arg Asp
100 105 110
His Thr Val Lys Glu Lys Leu Val Glu Arg Gly Ile Ser Val Gln Ser
115 120 125
Tyr Asn Gly Asp Leu Leu Tyr Glu Pro Trp Glu Ile Tyr Cys Glu Lys
130 135 140
Gly Lys Pro Phe Thr Ser Phe Asn Ser Tyr Trp Lys Lys Cys Leu Asp
145 150 155 160
Met Ser Ile Glu Ser Val Met Leu Pro Pro Pro Trp Arg Leu Met Pro
165 170 175
Ile Thr Ala Ala Ala Glu Ala Ile Trp Ala Cys Ser Ile Glu Glu Leu
180 185 190
Gly Leu Glu Asn Glu Ala Glu Lys Pro Ser Asn Ala Leu Leu Thr Arg
195 200 205
Ala Trp Ser Pro Gly Trp Ser Asn Ala Asp Lys Leu Leu Asn Glu Phe
210 215 220
Ile Glu Lys Gln Leu Ile Asp Tyr Ala Lys Asn Ser Lys Lys Val Val
225 230 235 240
Gly Asn Ser Thr Ser Leu Leu Ser Pro Tyr Leu His Phe Gly Glu Ile
245 250 255
Ser Val Arg His Val Phe Gln Cys Ala Arg Met Lys Gln Ile Ile Trp
260 265 270
Ala Arg Asp Lys Asn Ser Glu Gly Glu Glu Ser Ala Asp Leu Phe Leu
275 280 285
Arg Gly Ile Gly Leu Arg Glu Tyr Ser Arg Tyr Ile Cys Phe Asn Phe
290 295 300
Pro Phe Thr His Glu Gln Ser Leu Leu Ser His Leu Arg Phe Phe Pro
305 310 315 320
Trp Asp Ala Asp Val Asp Lys Phe Lys Ala Trp Arg Gln Gly Arg Thr
325 330 335
Gly Tyr Pro Leu Val Asp Ala Gly Met Arg Glu Leu Trp Ala Thr Gly
340 345 350
Trp Met His Asn Arg Ile Arg Val Ile Val Ser Ser Phe Ala Val Lys
355 360 365
Phe Leu Leu Leu Pro Trp Lys Trp Gly Met Lys Tyr Phe Trp Asp Thr
370 375 380
Leu Leu Asp Ala Asp Leu Glu Cys Asp Ile Leu Gly Trp Gln Tyr Ile
385 390 395 400
Ser Gly Ser Ile Pro Asp Gly His Glu Leu Asp Arg Leu Asp Asn Pro
405 410 415
Ala Leu Gln Gly Ala Lys Tyr Asp Pro Glu Gly Glu Tyr Ile Arg Gln
420 425 430
Trp Leu Pro Glu Leu Ala Arg Leu Pro Thr Glu Trp Ile His His Pro
435 440 445
Trp Asp Ala Pro Leu Thr Val Leu Lys Ala Ser Gly Val Glu Leu Gly
450 455 460
Thr Asn Tyr Ala Lys Pro Ile Val Asp Ile Asp Thr Ala Arg Glu Leu
465 470 475 480
Leu Ala Lys Ala Ile Ser Arg Thr Arg Glu Ala Gln Ile Met Ile Gly
485 490 495
Ala Ala Pro Asp Glu Ile Val Ala Asp Ser Phe Glu Ala Leu Gly Ala
500 505 510
Asn Thr Ile Lys Glu Pro Gly Leu Cys Pro Ser Val Ser Ser Asn Asp
515 520 525
Gln Gln Val Pro Ser Ala Val Arg Tyr Asn Gly Ser Lys Arg Val Lys
530 535 540
Pro Glu Glu Glu Glu Glu Arg Asp Met Lys Lys Ser Arg Gly Phe Asp
545 550 555 560
Glu Arg Glu Leu Phe Ser Thr Ala Glu Ser Ser Ser Ser Ser Ser Val
565 570 575
Phe Phe Val Ser Gln Ser Cys Ser Leu Ala Ser Glu Gly Lys Asn Leu
580 585 590
Glu Gly Ile Gln Asp Ser Ser Asp Gln Ile Thr Thr Ser Leu Gly Lys
595 600 605
Asn Gly Cys Lys
610
<210> 21
<211> 675
<212> PRT
<213> Arabidopsis thaliana
<400> 21
Met Glu Glu Ile Ser Thr Asp Pro Val Val Pro Ala Val Lys Pro Asp
1 5 10 15
Pro Arg Thr Ser Ser Val Gly Glu Gly Ala Asn Arg His Glu Asn Asp
20 25 30
Asp Gly Gly Ser Gly Gly Ser Glu Ile Gly Ala Pro Asp Leu Asp Lys
35 40 45
Asp Leu Leu Cys Pro Ile Cys Met Gln Ile Ile Lys Asp Ala Phe Leu
50 55 60
Thr Ala Cys Gly His Ser Phe Cys Tyr Met Cys Ile Ile Thr His Leu
65 70 75 80
Arg Asn Lys Ser Asp Cys Pro Cys Cys Ser Gln His Leu Thr Asn Asn
85 90 95
Gln Leu Tyr Pro Asn Phe Leu Leu Asp Lys Leu Leu Lys Lys Thr Ser
100 105 110
Ala Arg His Val Ser Lys Thr Ala Ser Pro Leu Asp Gln Phe Arg Glu
115 120 125
Ala Leu Gln Arg Gly Cys Asp Val Ser Ile Lys Glu Val Asp Asn Leu
130 135 140
Leu Thr Leu Leu Ala Glu Arg Lys Arg Lys Met Glu Gln Glu Glu Ala
145 150 155 160
Glu Arg Asn Met Gln Ile Leu Leu Asp Phe Leu His Cys Leu Arg Lys
165 170 175
Gln Lys Val Asp Glu Leu Asn Glu Val Gln Thr Asp Leu Gln Tyr Ile
180 185 190
Lys Glu Asp Ile Asn Ala Val Glu Arg His Arg Ile Asp Leu Tyr Arg
195 200 205
Ala Arg Asp Arg Tyr Ser Val Lys Leu Arg Met Leu Gly Asp Asp Pro
210 215 220
Ser Thr Arg Asn Ala Trp Pro His Glu Lys Asn Gln Ile Gly Phe Asn
225 230 235 240
Ser Asn Ser Leu Ser Ile Arg Gly Gly Asn Phe Val Gly Asn Tyr Gln
245 250 255
Asn Lys Lys Val Glu Gly Lys Ala Gln Gly Ser Ser His Gly Leu Pro
260 265 270
Lys Lys Asp Ala Leu Ser Gly Ser Asp Ser Gln Ser Leu Asn Gln Ser
275 280 285
Thr Val Ser Met Ala Arg Lys Lys Arg Ile His Ala Gln Phe Asn Asp
290 295 300
Leu Gln Glu Cys Tyr Leu Gln Lys Arg Arg Gln Leu Ala Asp Gln Pro
305 310 315 320
Asn Ser Lys Gln Glu Asn Asp Lys Ser Val Val Arg Arg Glu Gly Tyr
325 330 335
Ser Asn Gly Leu Ala Asp Phe Gln Ser Val Leu Thr Thr Phe Thr Arg
340 345 350
Tyr Ser Arg Leu Arg Val Ile Ala Glu Ile Arg His Gly Asp Ile Phe
355 360 365
His Ser Ala Asn Ile Val Ser Ser Ile Glu Phe Asp Arg Asp Asp Glu
370 375 380
Leu Phe Ala Thr Ala Gly Val Ser Arg Cys Ile Lys Val Phe Asp Phe
385 390 395 400
Ser Ser Val Val Asn Glu Pro Ala Asp Met Gln Cys Pro Ile Val Glu
405 410 415
Met Ser Thr Arg Ser Lys Leu Ser Cys Leu Ser Trp Asn Lys His Glu
420 425 430
Lys Asn His Ile Ala Ser Ser Asp Tyr Glu Gly Ile Val Thr Val Trp
435 440 445
Asp Val Thr Thr Arg Gln Ser Leu Met Glu Tyr Glu Glu His Glu Lys
450 455 460
Arg Ala Trp Ser Val Asp Phe Ser Arg Thr Glu Pro Ser Met Leu Val
465 470 475 480
Ser Gly Ser Asp Asp Cys Lys Val Lys Val Trp Cys Thr Arg Gln Glu
485 490 495
Ala Ser Val Ile Asn Ile Asp Met Lys Ala Asn Ile Cys Cys Val Lys
500 505 510
Tyr Asn Pro Gly Ser Ser Asn Tyr Ile Ala Val Gly Ser Ala Asp His
515 520 525
His Ile His Tyr Tyr Asp Leu Arg Asn Ile Ser Gln Pro Leu His Val
530 535 540
Phe Ser Gly His Lys Lys Ala Val Ser Tyr Val Lys Phe Leu Ser Asn
545 550 555 560
Asn Glu Leu Ala Ser Ala Ser Thr Asp Ser Thr Leu Arg Leu Trp Asp
565 570 575
Val Lys Asp Asn Leu Pro Val Arg Thr Phe Arg Gly His Thr Asn Glu
580 585 590
Lys Asn Phe Val Gly Leu Thr Val Asn Ser Glu Tyr Leu Ala Cys Gly
595 600 605
Ser Glu Thr Asn Glu Val Tyr Val Tyr His Lys Glu Ile Thr Arg Pro
610 615 620
Val Thr Ser His Arg Phe Gly Ser Pro Asp Met Asp Asp Ala Glu Glu
625 630 635 640
Glu Ala Gly Ser Tyr Phe Ile Ser Ala Val Cys Trp Lys Ser Asp Ser
645 650 655
Pro Thr Met Leu Thr Ala Asn Ser Gln Gly Thr Ile Lys Val Leu Val
660 665 670
Leu Ala Ala
675
<210> 22
<211> 514
<212> PRT
<213> Arabidopsis thaliana
<400> 22
Met Leu Leu His Gln His Gln Gln Pro Gln Gln Asp Glu Asn Met Ser
1 5 10 15
Asn Leu Thr Ser Ala Ser Gly Asp Gln Ala Ser Val Ser Ser Gly Asn
20 25 30
Ile Thr Glu Ala Ser Gly Ser Asn Tyr Phe Pro His His Gln Gln Gln
35 40 45
Gln Glu Gln Gln Gln Gln Gln Ile Gln Lys Leu Ser Cys Ser Trp Thr
50 55 60
Asp Ser Leu Phe Gln Leu Phe Asp Thr Val Thr Phe Leu Glu Ile Leu
65 70 75 80
Tyr Pro Glu Ser Glu Val Ile Ala Leu Ser Pro Lys Thr Leu Met Ala
85 90 95
Thr Asn Arg Phe Val Cys Glu Ile Cys Asn Lys Gly Phe Gln Arg Asp
100 105 110
Gln Asn Leu Gln Leu His Arg Arg Gly His Asn Leu Pro Trp Lys Leu
115 120 125
Lys Gln Arg Ser Asn Lys Glu Val Ile Arg Lys Lys Val Tyr Val Cys
130 135 140
Pro Glu Ala Ser Cys Val His His Asp Pro Ser Arg Ala Leu Gly Asp
145 150 155 160
Leu Thr Gly Ile Lys Lys His Phe Cys Arg Lys His Gly Glu Lys Lys
165 170 175
Trp Lys Cys Asp Lys Cys Ser Lys Lys Tyr Ala Val Gln Ser Asp Cys
180 185 190
Lys Ala His Ser Lys Thr Cys Gly Thr Lys Glu Tyr Arg Cys Asp Cys
195 200 205
Gly Thr Leu Phe Ser Arg Arg Asp Ser Phe Ile Thr His Arg Ala Phe
210 215 220
Cys Glu Ala Leu Ala Glu Glu Thr Ala Arg Glu Val Val Ile Pro Gln
225 230 235 240
Asn Gln Asn Asn Asn Gln Pro Asn Pro Leu Leu Ile His Gln Ser Ala
245 250 255
Ser His Pro His His His His Gln Thr Gln Pro Thr Ile Asn Val Ser
260 265 270
Ser Ser Ser Ser Ser Ser His Asn His Asn Ile Ile Asn Ser Leu His
275 280 285
Phe Asp Thr Asn Asn Gly Asn Thr Asn Asn Ser Asn Asn Ser Asn Asn
290 295 300
His Leu His Thr Phe Pro Met Lys Lys Glu Gln Gln Ser Asn Asp His
305 310 315 320
Ile Met Asn Tyr His His Ser Ile Ile Pro Pro Trp Leu Ala Pro Gln
325 330 335
Pro His Ala Leu Thr Ser Ser Asn Pro Asn Pro Ser Asn Gly Gly Gly
340 345 350
Gly Gly Gly Ser Leu Phe Ser Leu Ala Ser Pro Ala Met Ser Ala Thr
355 360 365
Ala Leu Leu Gln Lys Ala Ala Gln Met Gly Ser Thr Lys Thr Pro Pro
370 375 380
Leu Pro Pro Thr Thr Ala Tyr Glu Arg Ser Thr His Asn Asn Asn Leu
385 390 395 400
Thr Thr Thr Met Ala Ala Met Met Thr Ser Pro Ser Gly Phe Ile Ser
405 410 415
Ser Asn Asn Asn Asn His Val Leu Phe Gln Asp Tyr Asn Ala Ser Gly
420 425 430
Phe Asp Asn His Gly Arg Glu Glu Ala Phe Asp Asp Thr Phe Gly Gly
435 440 445
Phe Leu Arg Thr Asn Glu Val Thr Ala Ala Ala Gly Ser Glu Lys Ser
450 455 460
Thr Lys Ser Gly Gly Gly Glu Gly Leu Thr Arg Asp Phe Leu Gly Leu
465 470 475 480
Arg Pro Leu Met Ser His Asn Glu Ile Leu Ser Phe Ala Gly Leu Gly
485 490 495
Ser Cys Ile Asn Ser Ser Ala Ser Asp Gln Leu His Pro Lys Pro Trp
500 505 510
Gln Gly
Claims (13)
- 애기장대 유래 LUX (LUX ARRHYTHMO), IDD7 (INDETERMINATE(ID)-DOMAIN 7), SVP (SHORT VEGETATIVE PHASE), SEP3 (SEPALLATA 3), CALS1 (CALLOSE SYNTHASE 1), WRI1 (WRINKLED 1), KCS2 (3-KETOACYL-COA SYNTHASE 2), KCS20, CRY2 (CRYPTOCHROME 2) 또는 COP1 (CONSTITUTIVE PHOTOMORPHOGENIC 1) 단백질을 코딩하는 유전자의 발현을 식물세포에서 조절하는 단계를 포함하는, 식물체의 재분화(regeneration) 효율을 조절하는 방법.
- 제1항에 있어서, 상기 애기장대 유래 LUX, IDD7, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 및 COP1 단백질은 각각 순서대로 서열번호 12 내지 21의 아미노산 서열로 이루어진 것을 특징으로 하는 식물체의 재분화 효율을 조절하는 방법.
- 제1항에 있어서, 상기 애기장대 유래 LUX, IDD7, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질을 코딩하는 유전자의 발현을 식물세포에서 저해시켜 야생형 식물체에 비해 식물체의 재분화 효율을 증가시키는 것을 특징으로 하는, 식물체의 재분화 효율을 조절하는 방법.
- 제3항에 있어서, 상기 IDD7 단백질을 코딩하는 유전자의 발현 저해는 서열번호 22의 아미노산 서열로 이루어진 애기장대 유래 IDD11 단백질을 코딩하는 유전자의 발현 저해를 추가로 포함하는 것을 특징으로 하는, 식물체의 재분화 효율을 조절하는 방법.
- 제3항에 있어서, 상기 애기장대 유래 유전자의 발현저해는 VIGS (Virus-induced gene silencing) 벡터, RNAi (RNA interference) 또는 안티센스 RNA, T-DNA 삽입, 내생 트랜스포존(transposon), 방사선 조사 또는 유전자 교정 시스템을 통한 돌연변이 유발을 이용하여 발현을 저해하는 것을 특징으로 하는, 식물체의 재분화 효율을 조절하는 방법.
- 제1항에 있어서, 상기 재분화는 지상부 재분화인 것을 특징으로 하는, 식물체의 재분화 효율을 조절하는 방법.
- 애기장대 유래 LUX, IDD7, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질을 코딩하는 유전자를 포함하는 재조합 벡터로 식물세포를 형질전환하는 단계를 포함하는, 재분화 효율이 조절된 형질전환 식물체의 제조방법.
- 제7항에 있어서, 상기 애기장대 유래 LUX, IDD7, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질을 코딩하는 유전자의 발현을 식물세포에서 저해시켜 야생형에 비해 식물체의 재분화 효율이 증가된 것을 특징으로 하는 형질전환 식물체의 제조방법.
- 제8항에 있어서, 상기 IDD7 단백질을 코딩하는 유전자의 발현 저해는 IDD11 단백질을 코딩하는 유전자의 발현 저해를 추가로 포함하는 것을 특징으로 하는 형질전환 식물체의 제조방법.
- 제7항 내지 제9항 중 어느 한 항의 방법에 의해 제조된 재분화 효율이 조절된 형질전환 식물체.
- 제10항에 따른 식물체의 형질전환된 종자.
- 애기장대 유래 LUX, IDD7/11, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질을 코딩하는 유전자를 유효성분으로 포함하는, 식물체의 재분화 효율 조절용 조성물.
- 애기장대 유래 LUX, IDD7/11, SVP, SEP3, CALS1, WRI1, KCS2, KCS20, CRY2 또는 COP1 단백질을 코딩하는 유전자의 발현이 저해되어, 야생형에 비해 재분화 효율이 증가된 돌연변이 식물체.
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