KR960705209A - 녹색 형광 단백의 용도 - Google Patents

녹색 형광 단백의 용도

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KR960705209A
KR960705209A KR1019960701205A KR19960701205A KR960705209A KR 960705209 A KR960705209 A KR 960705209A KR 1019960701205 A KR1019960701205 A KR 1019960701205A KR 19960701205 A KR19960701205 A KR 19960701205A KR 960705209 A KR960705209 A KR 960705209A
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cell
protein
green fluorescent
cells
fluorescent protein
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마틴 찰피
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잭 엠. 그랜노위츠
더 트러스티스 오브 콜롬비아 유니버시티 인 더 시티 오브 뉴욕
원본미기재
우드즈 홀 오션오그래픽 인스티튜션
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Priority claimed from US08/119,678 external-priority patent/US5491084A/en
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Publication of KR960705209A publication Critical patent/KR960705209A/ko

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Abstract

본 발명은 녹색 형광 단백질을 코딩하는 DNA 서열에 작용가능하게 연결된, 녹색 형광 단백질을 코딩하는 유전자 이외의 유전자로부터의 조절요소를 가진 DNA 분자를 포함하는 세포를 제공한다. 본 발명은 또한 상기 세포를 포함하는 생물체를 제공한다.
본 발명은 관심있는 단백질을 발현하는 세포를 선택하는 방법을 제공한다. 이 방법은 a) 관심있는 단백질을 코딩하는 DNA 서열을 가진 DNA I 분자와 녹색 형광 단백질을 코딩하는 DNA 서열을 가진 DNAⅡ 분자를 세포내에 도입하고; b) 도입된 세포를 관심있는 단백질 및 녹색 형광 단백질의 발현을 허용하는 조건하에서 배양하고; 그리고 c) 녹색 형광 단백질을 발현하는 배양된 세포를 선택함으로써, 관심있는 단백질을 발현하는 세포를 선택하는 것을 포함하는 단계를 포함한다. 마지막으로, 본 발명은 녹색 형광 단백질의 다양한 용도를 제공한다.

Description

녹색 형광 단백의 용도
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음

Claims (36)

  1. 녹색 형광 단백질을 코딩하는 DNA 서열에 작용가능하게 연결된, 녹색 형광 단백질을 코딩하는 유전자 이외의유전자로부터의 조절요소를 가진 DNA 분자를 포함하는 세포.
  2. 제1항에 있어서, 사기 세포가 실질상 세균세포, 효모세포, 진균세포, 곤충세포, 선충세포, 식물 또는 동물세포로 이루어지는 군으로부터 선택되는 것을 특징으로 하는 세포.
  3. 제1항에 있어서, 상기 조절요소가 프로모터인 것을 특징으로 하는 세포.
  4. 제3항에 있어서, 상기 프로모터가 중금속에 의해 활성화되는 것을 특징으로 하는 세포.
  5. 제3항에 있어서, 상기 프로모터가 P450 유전자로부터 유래하는 것을 특징으로 하는 세포.
  6. 제3항에 있어서, 상기 프로모터가 스트레스 단백질을 코딩하는 유전자로부터 유래함을 특징으로 하는 세포.
  7. 제6항에 있어서, 상기 스트레스 단백질이 열-쇼크 단백질인 것을 특징으로 하는 세포.
  8. 제3항에 있어서, 상기 프로모터가 세포 생존에 필요한 유전자로부터 유래하는 것을 특징으로 하는 세포.
  9. 제1항에 있어서, 상기 조절요소가 인핸서인 것을 특징으로 하는 세포.
  10. 제1항에 있어서, 상기 DNA 서열이 Aequorea Victoria 녹색 형광 단백질을 코딩하는 것을 특징으로 하는 세포.
  11. 제1항에 있어서, 상기 세포가 E. coil 세포인 것을 특징으로 하는 세포.
  12. 제11항에 있어서, 상기 E. coil 세포가 SMC1(ATCC 수탁번호 제69554호)로 명명된 것임을 특징으로 하는 세포.
  13. 제1항의 세포로부터 분리된 녹색 형광 단백질.
  14. 제1항의 세포를 포함하는 생물체.
  15. 제14항에 있어서, 상기 생물체가 Caenorhabditis elegans 인 것을 특징으로 하는 생명체.
  16. 제15항에 있어서, 상기 조절요소가 스트레스 단백질에 대한 것임을 특징으로 하는 생물체.
  17. 제16항에 있어서, 상기 스트레스 단백질이 열-쇼크 단백질인 것을 특징으로 하는 생물체.
  18. a) 제1항의 세포를 배양하고; 그리고 b) 상기 세포로부터 생성된 녹색 형광 단백질을 분리 및 정제하는 것을 포함하는 녹색 형광 단백질을 생산하는 방법.
  19. 제18항에 있어서, 상기 세포가 E. coil 세포인 것을 특징으로 하는 방법.
  20. 제19항에 있어서, 상기 세포가 호기적으로 배양되는 것을 특징으로 하는 방법.
  21. 제18항에 있어서, 상기 세포가 SMC1(ATCC 수탁번호 제69554호로 명명된 것임을 특징으로 하는 세포.
  22. a) 관심있는 단백질을 코딩하는 DNA 서열을 가진 DNA I 분자의 녹색 형광 단백질을 코딩하는 DNA 서열을 가진 DNAⅡ 분자를 세포내에 도입하고; b) 도입된 세포를 관심있는 단백질 및 녹색 형광 단백질의 발현을 허용하는 조건하에서 배양하고; 그리고 c) 녹색 형광 단백질을 발현하는 배양된 세포를 선택함으로써, 관심있는 단백질을 발현하는 세포를 선택하는 것을 포함하는, 관심있는 단백질을 발현하는 세포를 선택하는 방법.
  23. 제22항에 있어서, DNA I과 DNAⅡ가 연결된 것을 특징으로 하는 방법.
  24. 제22항에 있어서, 상기 세포가 실질상 세균세포, 효모세포, 진균세포, 곤충세포, 선충세포, 식물 또는 동물세포로 이루어지는 군으로부터 선택되는 것을 특징으로 하는 방법.
  25. 제22항에 있어서, 상기 DNAⅡ가 Aequorea Victoria 녹색 형광 단백질을 암호화하는 것을 특징으로 하는 방법.
  26. a) 녹색 형광 단백질을 코딩하는 DNA 서열에 연결된 관심있는 단백질을 코딩하는 DNA 서열을 가진 DNA 분자를 이 DNA 분자에 의해 생산되는 단백질이 녹색 형광 단백질에 융합된 관심있는 단백질을 갖도록 세포내로 도입하고; b) 융합된 단백질의 발현을 허용하는 조건하에서 세포를 배양하고; 그리고 c) 상기 세포내에 녹색 형광 단백질로 이루어진 융합된 단백질의 위치를 탐지함으로써, 세포내에 관심있는 단백질의 위치를 확인하는 단계를 포함하는, 세포내에서 관심있는 단백질의 위치 확인 방법.
  27. 제26항에 있어서, 상기 세포가 관심있는 단백질을 정상적으로 발현하는 것을 특징으로 하는 방법.
  28. 제26항에 있어서, 상기 DNA가 Aequorea Victoria 녹색 형광 단백질을 코딩하는 것을 특징으로 하는 방법.
  29. 하기 단계들을 포함하는 세포내에서 우전자의 발현을탐지하는 방법: a) 녹색 형광 단백질을 코딩하는 DNA 서열에 연결된 상기 유전자의 DNA 서열을 가진 DNA 분자를 상기 유전자의 조절요소가 녹색 형광 단백질의 발현을 제어하도록 세포내에 도입하고; b) 상기 세포를 사기 유전자의 발현을 허용하는 조건하에서 배양하고; 그리고 c) 상기 세포내에서 녹색 형광 단백질의 발현을 탐지함으로써, 상기 세포내에서 상기 유전자의 발현을 보이는 단계.
  30. 하기 단계들을 포함하는 피검체에서 유전자의 발현을 탐지하는 방법: a) 녹색 형광 단백질을 코딩하는 DNA 서열에 연결된 상기 유전자의 DNA 서열을 가진 DNA 분자를 상기 우전자의 조절요소가 녹색 형광 단백질의 발현을 제어하도록 DNA 분자를 피검체의 세포내에 도입하고: b) 상기 융합된 단백질의 발현을 허용하는 조건하에서 상기 세포를 배양하고; 그리고 c) 상기 세포내에서 녹색 형광 단백질의 발현을 탐지함으로써, 상기 세포내에서 상기 유전자의 발현을 지시하는 단계.
  31. 제29항에 또는 제30항에 있어서, 상기 DNA가 Aequorea Victoria 녹색 형광 단백질을 코딩하는 것을 특징으로 하는 방법.
  32. 하기 단계들을 포함하는, 피검체에서 DNA 서열의전사의 조직-특이성을 결정하는 방법: a) 녹색 형광 단백질을 코딩하는 DNA 서열에 연결된 DNA 서열을 가진 DNA 분자를 피검체내에서 상기 DNA 서열 유전자가 녹색 형광 단백질의 발현을 제어하도록 세포내에 도입하고; b) 녹색 형광 단백질의 발현을 허용하는 조건하에서 피검체를 배양하고; 그리고 c) 피검체의 상인한 조직들에서 녹색 형광 단백질의 발현을 탐지함으로써, 상기 DNA 서열의 조직-특이성을 결정하는 단계.
  33. a) 용액내에서 제4항 또는 제5항의 세포를 배양하고; 그리고 b) 중금속의 존재를 나타내는 녹색 형광 단백질의 발현을 탐지하는 단계를 포함하는 용액내에서 중금속을 탐지하는 방법.
  34. a) 용액내에서제4,5,6 또는 7항의 세포를 배양하고; 그리고 b) 오염물질의 존재를 나타내는 녹색 단백질의 발현을 탐지하는 단계를 포함하는 용액내에서 오염물질을 탐지하는 방법.
  35. a) 녹색 형광 단백질을 코딩하는 DNA 분자를 같은 해독틀로 기지 아미노산 서열을 암호화하는 DNA 분자와 연결하고; b) 단계 a)의 연결된 DNA 분자에 의해 코딩되는 형광 단백질의 발현을 허용하는 발현 시스템에 연결된 DNA 분자를 도입하고; 그리고 c) 단계 b)의 발현된 형광 단백질의 분량을 결정함으로써, 형광 단백질 분자량 마커를 생산하는 단계.
  36. 제35항에 있어서, 상기의 발현된 단백질의 정체를 추가로 포함하는 방법.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019960701205A 1993-09-10 1994-09-09 녹색 형광 단백의 용도 KR960705209A (ko)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US08/119,678 US5491084A (en) 1993-09-10 1993-09-10 Uses of green-fluorescent protein
US19227494A 1994-02-04 1994-02-04
US08/119,678 1994-02-04
US08/192,274 1994-02-04
PCT/US1994/010165 WO1995007463A1 (en) 1993-09-10 1994-09-09 Uses of green fluorescent protein

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