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KR900700601A - 레티노 산 수용체 조성물 및 방법 - Google Patents

레티노 산 수용체 조성물 및 방법

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KR900700601A
KR900700601A KR1019890701441A KR890701441A KR900700601A KR 900700601 A KR900700601 A KR 900700601A KR 1019890701441 A KR1019890701441 A KR 1019890701441A KR 890701441 A KR890701441 A KR 890701441A KR 900700601 A KR900700601 A KR 900700601A
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마아크 에반스 로날드
기구어 빈센트
세바스챤 옹 에스테리타
세라노 세귀 프루디마르
우메소노 카주히코
캐롤린 톰프슨 카테린
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델버어트 어네스트 글렌쯔
더 솔크 인스티튜트 포오 바이오로지칼 스터디이즈
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Abstract

내용 없음

Description

레티노 산 수용체 조성물 및 방법
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음

Claims (29)

  1. 센스(sense) 스트랜드가 레티노 산 수용체 단백질의 특징적인 리간드-결합 및 전사-활성화 성질을 갖는 단백질의 일차적 서열에 대해 암호화하는 트리플리트(triplet)의 서열을 갖는 단편으로 구성되어 있고, 상기 레티노 산 수용체 단백질로 발현될 수 있는 본질적으로 순수한 이중-스트랜드 DNA.
  2. 제1항에 있어서, 상기 단편내 트리플리트의 서열이 제1B-1도, 제1B-2도 및 제1B-3도에서 보여주는 아미노산 서열과 유사한 본질적인 아미노산을 갖는 단백질의 일차적 서열에 대해 암호화하는 본질적으로 순수한 DNA.
  3. 제1B도, 제1B-2도 및 제1B-3도에서 보여준 DNA서열과 상당한 서열 유사성을 갖는 본질적으로 순수한 DNA서열.
  4. 제1항에 있어서, 단편내 트리플리트의 서열이 인간의 레티노산 수용체 단백질의 특징적인 라간드-결합 및 전사-활성화 성질을 갖는 단백질의 일차적 서열에 대해 암호화하는 본질적으로 순수한 DNA.
  5. 플라스미드 phRAR1(ATCC#40392).
  6. 플라스미드 phRAR1(ATCC#40392)의 DNA와 상당한 서열 유사성을 갖는 본질적으로 순수한 DNA 서열.
  7. (a) 번역 시작 코돈 트리플리트의 다운스트림(downstream) 및 상기 코돈에 상당하는 트리플리트를 갖는 프레임안에 있는, 번역 중단 코돈에 상당하는 하나의 트리플리트, 및 (b) 가장 긴 오픈리딩(open reading)프레임으로서, 테리노 산 수용체 단백질의 일차적 서열에 대해 암호화하는 트리플리트의 서열을 갖는 본질적으로 순수한 단편을 갖는 본질적으로 순수한 DNA단편.
  8. 인간의 레티노 산 수용체를 암호화하는 본직적으로 순수한 DNA.
  9. 제1항 내지 제8항중 어느 한항에 있어서 청구된 임의 DNA와 상당한 서열 유사성을 갖는 본질적으로 순수한 DNA.
  10. 제1항 내지 제8항중 어느 한항에 청구된 임의 DNA의 돌연변이체.
  11. 제1항 내지 제8항중 어느 한항에 청구된 임의 본질적으로 순수한 DNA서열로부터 전사된 본질적으로 순수한 mRNA.
  12. 단백질의 아미노산 서열이 제1B-1도, 제1B-2도 및 제1B-3도에서 보여준 아미노산 서열과 유사한 적어도 약 50% 아미노산을 갖는 본질적으로 순수한 단백질.
  13. 제1항 내지 제8항중 어느 한항에 청구된 임의 본질적으로 순수한 DNA서열에 의해 암호화된 본질적으로 순수한 단백질.
  14. 제1항 내지 제8항중 어느 한항에 청구된 본질적으로 순수한 임의의 DNA에 의해 형질전환된 세포.
  15. N-말단 영역, DNA-결합 영역 및 리간드-결합 영역이 GR, MR, TR, ERR 및 RR로 이루어진 모(parental) 수용체들의 일반 군으로부터 선택된 공지의 수용체로부터 유래되고, 키메라가(1) hGR, hMR, hERR1, hERR2, rTRα, hT3α, hT3β, hRARα및 hRARβ로 이루어진 모 수용체의 군으로부터 선택된 N-말단 영역, (2) hGR, hMR, hERR1, hERR2, rTRα, hT3α, hT3β, hRARα및 hRARβ로 이루어진 모 수용체의 군으로부터 선택된 DNA-결합 영역, 및 (3) hGR, hMR, hERR1, hERR2, rTRα, hT3α, hT3β, hRARα및 hRARβ로 이루어진 모 수용체의 군으로부터 선택된 리간드-결합영역을 가지며, 임의 하나의 키메릭 수용체가 적어도 두 개의 다른 "모" 급원으로부터 유래되는 N-말단 영역, DNA-결합영역, 및 리간드-결합 영역을 가지므로 결코 임의 야생-형 수용체와는 동일하지 않은, N-말단 영역, DNA-결합영역, 및 리간드-결합 영역을 갖는 키메릭 수용체.
  16. GRR, GRG GGR, RGG, RGR, RRG, TTG, GTT, GTG, GGT, TGG, TGT, TTR, TRT, TRR, RTT, RTR, RRT, GTT, GTG, GGT, TGG, TGT, 및 TTG로 이루어진 군으로부터 선택된 키메릭 수용체를 암호화하는 DNA 서열.
  17. 임의 주어진 세포내에 외생의 백그라운드 결합 또는 전사적 활성화 작용 수준을 초과하는 활성을 갖거나, 상당하는 자연적으로 생긴 수용체 DNA-결합 영역의 DNA-결합 또는 전사-활성화 작용의 적어도 약 5% 및/또는 상당하는 자연적으로 생긴 리간드-결합영역의 리간드-결합 활성의 약 5%를 가질, 그런 키메릭 수용체 암호화 DNA로부터 전사된 mRNA의 번역 또는 본 발명의 키메릭 DNA의 발현에 의해 만들어진 키메릭 수용체 단백질.
  18. (a) 추정상 리간드-결합 영역 및 추정상 DNA-결합 영역을 갖는 DNA서열을 분리시키고; (b) 단계(a)의 DNA 서열의 DNA-결합 영역 부분을 공지된 리간드-반응성 수용체 단백질로부터 DNA-결합 영역 부분으로 대치시킴으로써 키메릭 유전자를 구성하고; (c) (1) (b) 단계로 부터의 키메릭 유전자, 및 (2) 호르몬 반응 요수가 단계 (b)의 키메릭 유전자에 의해 암호화된 수용체 단백질의 DNA-결합 영역 부분에 의해 활성화 될 수 있는 조작적 호르몬 반응성 요소에 기능적으로 연결된 리포터 유전자로 구성된 적합한 수용체-결핍 숙주 세포내로 형질전환시키고; (d) 단계(c)로부터 형질전환된 숙주 세포를 단계(b)의 키메릭 유전자에 의해 암호화된 키메릭 수용체 단백질의 리간드-결합 영역 부분과 잠재적으로 결합할 수 있는 일련의 후보 리간드와 경쟁시키고; (e) 리포터 유전자에 의해 암호화된 단백질의 수준의 변화에 의해 리포터 유전자의 유도를 조사하고; (f) 리포터 유전자의 단백질 생성물의 생산을 유도할 수 있는 리간드(들)를 리간드(들)로서 선택하는 것으로 구성되는, 수용체 단백질에 대한 기능 리간드를 확인하는 방법.
  19. 제18항 (b)에 있어서, 공지된 리간드-반응성 수용체 단백질이 글루코코리티코이드 수용체, 미네랄로코르티코이드 수용체, 인간의 티로이드 수용체 알파 및 베타 및 쥐의 티로이드 수용체 알파, 에스트로겐-관련 수용체 hERR1 및 hERR2 및 레티노 산 수용체 알파 및 베타로 이루어진 군으로부터 선택되는 방법.
  20. 제18항 (c)에 있어서, 숙주 세포가 COS세포인 방법.
  21. 제18항 (c) (2)에 있어서, 리포터 유전자가 클로람페니콜 아세틸트랜스퍼라제(CAT) 및 개똥벌레 루시페라제 유전자로 이루어진 군으로부터 선택되는 방법.
  22. 제18항 (c) (2)에 있어서, 호르몬 반응 요소가 야생-형, 조작 또는 합성(1) 글루코코르티코이드 반응 요소, (2) 트로이드 반응 요소, (3) 미네랄로코르티코이드 반응 요소, (4) 에스트로겐-관련 반응 요소, (5) 레티노 산 반응 요소, 및 (6) 비타민 D3반응 요소로 이루어진 군으로부터 선택되는 방법.
  23. 제22항에 있어서, 글루코코르티코이드 반응 요소가 유암비루스의 긴 말단 반복서열(MTV LTR)의 일부이고, 티로이드 반응 요소가 성장 호르몬 프로모터 서열의 일부인 방법.
  24. 제18항에 있어서, 단계 (f)에서 확인된 리간드(들)가 단계(f)에서 확인된 리간드(들) 및 단계(a)의 DNA 서열의 발현 생성물의 결합 성질을 평가함으로써 확증되는 방법.
  25. 세포가 (a) 두 번째 수용체 서열의 리간드-결합 영역의 조작부분(B)에 연결된 첫 번째 수용체 서열의 DNA-결합 영역의 조작부분(A)으로 구성된 키메릭 DNA서열(c) 및 (b) 첫 번째 수용체 서열의 DNA-결합 영역의 조작 부분이 리포터 서열에 기능적으로 연결되는 호르몬 반응 요소에 기능적으로 결합하여 활성화될 수 있는 조작적 호르몬 반응 요소에 기능적으로 연결된 리포터 헥산 서열을 함유하는 세포내 수용체 단백질에 대한 기능 리간드를 확인하고, 세포를 적어도 하나의 후보 리간드와 경쟁시킨 다음 리포터 서열의 발현 생성물의 양으로 변화에 의한 리포터 헥산 서열의 유도를 조사하는 것으로 구성되는 방법.
  26. 제25항에 있어서, 상기 세포가 COS세포인 방법.
  27. 제25항에 있어서, 리포터 유전자가 클로람페니콜 아세틸 트랜스퍼라제(CAT)유전자 및 개똥벌레 루시페라제 유전자로 이루어진 군으로부터 선택되는 방법.
  28. 제25항에 있어서, 호르몬 반응 요소가 야생-형, 조작 또는 합성 (1) 글루코코르티코이드 반응 요소, (2) 티로이드 반응 요소, (3) 미네랄로코르티코이드 반응 요소, (4) 에스트로겐-관련 반응 요소, (5) 테티노산 반응 요소, 및 (6) 비타민 D3반응 요소로 이루어진 군으로부터 선택되는 방법.
  29. 제28항에 있어서, 글루코크르티코이드 반응 요소가 유방암 비루스의 긴 말단 반복 서열(MTV LTR)의 일부이고, 티로이드 반응 요소가 성장 호르몬 프로모터 서열의 일부인 방법.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019890701441A 1987-12-02 1988-12-01 레티노 산 수용체 조성물 및 이의 제조 방법 KR970009951B1 (ko)

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