KR880005270A - 단백질c의 발현 - Google Patents

단백질c의 발현 Download PDF

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KR880005270A
KR880005270A KR870011997A KR870011997A KR880005270A KR 880005270 A KR880005270 A KR 880005270A KR 870011997 A KR870011997 A KR 870011997A KR 870011997 A KR870011997 A KR 870011997A KR 880005270 A KR880005270 A KR 880005270A
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protein
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dna sequence
arg
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씨. 포스터 도날드
제이. 머레이 마크
엘. 버크너 캐틀린
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원본미기재
자이모 제네틱스. 인코포레이티드
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Abstract

내용 없음

Description

단백질C의 발현
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제1도는 PHC λ 6L 중의 단백질 C cDNA의 부분적인 제한지도이다.
제2도는 완전한 단백질 C cDNA의 뉴클레오티드 서열 및 단백질 C의 추측된아미노산 서열을 설명한 것이다.
제3도는 인체 단백질 C를 코드화하는 게놈 DNA의 제한효소지도이다.

Claims (15)

  1. 사람의 단백질 C 또는 사람의 활성화된 단백질 C 와 거의 동일한 생물학적 활성을 갖는 단백질을 코드화하고, 또한 고분자량쇄와 저분자량쇄 사이의 절단위치에 존재하는 아미노상 서열 (R1)n-R2-R3-R4(여기에서, R1, R2, R3및 R4은 Lys또는 Arg 이고 : n은 0, 1, 2 또는 3이다)을 코드화하는 DNA서열.
  2. 제1항에 있어서, 고분자량쇄와 저분자량쇄 사이의 절단위치에 존재하는 언급한 아미노상 서열이 Arg-Arg-Lys-Arg인 DNA 서열.
  3. 사람의 단백질 C 또는 사람의 활성화된 단백질 C와 거의 동일한 생물학적 활성을 갖는 단백질을 코드화하고, 또한 인자 VII, 인자 IX, 인자 X 프로트롬빈 및 단백질 S중에서 선택되는 단백질의 프리-프로 펩타이드를 코드화 하는 DNA서열.
  4. 사람의 단백질 C 또는 사람의 활성화된 단백질 C 와 거의 동일한 생물학적 활성을 가지며, 또한 잔기 158이 Ala, Ser, Thr및 Gly중에서 선택되는 비산성 아미노산잔기로 치환된 단백질을 코드화하는 DNA 서열.
  5. 사람의 단백질 C 또는 사람의 활성화된 단백질 C와 거의 동일한 생물학적 활성을 가지며, 잔기 154가 Lys 또는 Arg 중에서 선택되는 염기성 아미노산 잔기로 치환된 단백질을 코드화하는 DNA서열.
  6. 사람의 단백질 C 또는 사람의 활성화된 단백질 C와 거의 동일한 생물학적 활성을 가지며, 잔기 156내지 157의 Lys-Arg이 Lys-Arg 또는Arg- Arg 로 치환된 단백질을 코드화하는 DNA 서열.
  7. 제1항 내지 6항중 어느 한 항에 따른 DNA서열(여기에서, DNA서열은 폴리아데닐화 신호의 상부에 존재한다)의 상부에 존재하면서 DNA 서열의 전사를 유도하는 프로모터를 함유하는, 포유동물 숙주 세포 DNA에 도입시킬수 있는 발현벡터.
  8. 제7항에 따른 발현벡터로 형질전환된 포유동물 세포.
  9. 제8항에 있어서, 상기의 세포가, COS, BHK,쥐의 HepI, 쥐의 HepⅡ, TCMK,사람의 폐, 사람의 간종양, Hep G2, 마우스의 간, DUKX 및 293 세포중에서 선택되는 세포.
  10. 포유동물 숙주세포에 제7항에 따른 발현벡터를 도입시킨 다음, 적절한 배지에서 성장시키고, 이어서 상기의 발현벡터에 의해 코드화 되고 상기의 포유동물 숙주세포에 의해 생성된 단백질 생성물을 분리시킴을 특징으로 하여, 활성화된 경우에 사람의 활성화된 단백질 C와 거의 동일한 구조 및/ 또는 생물학적 활성을 갖는 단백질을 제조하는 방법.
  11. 제10항에 있어서, 상기의 세포가 COS, BHK, 쥐의 HepI, 쥐의 HepⅡ, TCMK, 사람의 폐, 사람의 간종양, Hep G2, 마우스 간, DUKX 및 293 세포중에서 선택되는 방법.
  12. BHK숙주 세포 DNA에 도입시킬 수 있으며, 제1항, 제5항 또는 제6항중 어느 한 항에 따른 DNA 서열의 상부에 DNA서열의 전사를 유도하는 프로모터가 존재하고, 상기 DNA 서열의 하부에 폴리아데닐화 신호가 존재하는 발현벡터를 BHK 숙주 세포에 도입시긴 다음 : 적절한 배지에 성장시키고 : 이어서 상기의 DNA서열에 의해 코드화 되고 상기의 BHK 숙주세포에 의해 생성된 단백질 생성물을 분리시킴을 특징으로 하여, 활성화된 경우에 사람의 활성화된 단백질 C와 거의 동일한 구조 및/ 또는 생물학적 활성을 갖는 단백질을 제조하는 방법.
  13. 제10항 내지 제12항중 어느 한 항에 있어서, 숙주 세포에 상기의 발현벡터와 함께 선별표지를 더 도입시키는 방법.
  14. 제10항 내지 제12항중 어느 한 항에 있어서, 사람의 활성화된 단백질의 C와 거의 동일한 생물학적 활성을 갖는 단백질을 제조하기 위하여 단백질 생성물을 활성화 시키는 단계를 더 포함하는 방법
  15. 제14항에 있어서, 활성화 단계가 단백질 생성물을 α-트룸빈, 트립신 및 독성 활성제중에서 선택되는 푸로테아제로 절단시키을 특징으로 하는 방법.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019870011997A 1986-10-29 1987-10-29 단백질 c를 암호화하는 dna 서열, 이를 함유하는 재조합 벡터, 재조합 벡터를 함유하는 포유동물 세포 및 단백질 c를 제조하는 방법 KR970002166B1 (ko)

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US06/924462 1986-10-29
US06/924,462 US4959318A (en) 1985-06-27 1986-10-29 Expression of protein C
US924462 1986-10-29

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KR880005270A true KR880005270A (ko) 1988-06-28
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EP (1) EP0266190B1 (ko)
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AT (1) ATE90966T1 (ko)
CA (1) CA1340263C (ko)
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EP0266190A3 (en) 1989-10-25
EP0266190B1 (en) 1993-06-23
ATE90966T1 (de) 1993-07-15
DK567087D0 (da) 1987-10-29
DE3786321T2 (de) 1993-09-30
NO874495D0 (no) 1987-10-28
CA1340263C (en) 1998-12-15
NO173741C (no) 1994-01-26
DK567087A (da) 1988-04-30
DE3786321D1 (de) 1993-07-29
NO173741B (no) 1993-10-18
JPH08252094A (ja) 1996-10-01
DK175183B1 (da) 2004-06-28
KR970002166B1 (ko) 1997-02-24
JP2922461B2 (ja) 1999-07-26
JPS6485084A (en) 1989-03-30
EP0266190A2 (en) 1988-05-04
JP2561677B2 (ja) 1996-12-11
NO874495L (no) 1988-05-02
US4959318A (en) 1990-09-25

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