KR950702635A - 감쇠 박테리아에서 재조합 융합 단백질의 발현(Expression of recombinant fusion proteins in attenuated bacteria) - Google Patents

감쇠 박테리아에서 재조합 융합 단백질의 발현(Expression of recombinant fusion proteins in attenuated bacteria) Download PDF

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KR950702635A
KR950702635A KR1019950700271A KR19950700271A KR950702635A KR 950702635 A KR950702635 A KR 950702635A KR 1019950700271 A KR1019950700271 A KR 1019950700271A KR 19950700271 A KR19950700271 A KR 19950700271A KR 950702635 A KR950702635 A KR 950702635A
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모하메드 아잠 칸
카를로스 에스테니오 호르마쉐
베르나르도 빌라레알-라모스
스티븐 네질 체트필드
고르돈 도우간
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크리스틴 헬렌 수덴
메데바 홀딩스 비. 브이
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Abstract

본 발명은 힌지부분에 의해 연결된 제1과 제2단백질로 인코드된 DNA 서열에 연결된 프로모터로 구성된 DNA 분자로 이때 프로모터는 주변환경 변화에 따라 반응하여 유도되는 활성을 가지고, 제1단백질은 파상풍 독소단편 C 또는 이의 에피토프가 된다. 본 발명은 제1항원서열, 힌지부분 그리고 3'단부에 제2항원 서열을 인코드하는 서열에 연결된 프로모터를 가지는 중간 단백질을 제공한다. 또한 본 발명은 DNA 융합단백질을 발현시키는 복제가능한 발현벡터, 이 벡터에 의해 저환된 박테리아와 박테리아에서 백신으로 사용되는 것을 제공한다.

Description

감쇠 박테리아에서 재조합 융합 단백질의 발현(Expression of recombinant fusion proteins in attenuated bacteria)
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제1도는 본 발명에 따른 중간 플라스미드 pTECH1의 구조를 설명한다.
제2도는 제2중간 플라스미드 pTECH2의 구조를 설명한다.
제3도는 시발물질로써 도면2의 중간 플라스미드를 이용하여 본 발명의 플라스미드의 구조를 설명한다.
제3도에서 B=BamHI, E=EcoRV, H=HindIII; X=Xbai; S=SpeI.

Claims (24)

  1. 힌지부분에 의해 연결된 제1과 제2단백질을 인코드하는 DNA 서열에 작동식으로 연결된 프로모터 서열로 구성된 DNA 구조에 있어서, 프로모터 서열은 주변환경에 따라 변화하여 유도될 수 있는 활성을 가지는 것을 특징으로 하는 DNA 구조.
  2. 제1항에 있어서, 단백질은 폴리펩티드 이류노겐인 것을 특징으로 하는 DNA 구조.
  3. 제1항에 있어서, 제1이형 단백질은 파상풍 독소단편 C 또는 이의 에피토프로 구성된 항원서열인 것을 특징으로 하는 DNA 구조.
  4. 제1과 제2이형 단백질에 연결된 DNA 서열에 작동식으로 연결된 프로모터 서열로 구성된 DNA 구조에 있어서, 제1이형 단백질은 파상풍 독소 단편 C 또는 이의 하나이상의 에피토프로 구성된 항원서열인 것을 특징으로 하는 DNA 구조.
  5. 제4항에 있어서, 프로모터 서열은 주변환경 변화에 반응하여 유도되는 하나의 활성을 가지는 것을 특징으로 하는 DNA 구조.
  6. 전술한 어느 한항에 있어서, 제1과 제2단백질은 이형인 것을 특징으로 하는 DNA 구조.
  7. 제4항 또는 5항에 있어서, 제1과 제2단백질은 힌지부분에 의해 연결되고, 힌지는 각 단백질과는 명백히 상이한 것을 특징으로 하는 DNA 구조.
  8. 진술한 어느 한항에 있어서, 프로모터 서열은 혐기성 조건에 의해 유도된 활성을 가지는 것을 특징으로 하는 DNA 구조.
  9. 제7항에 있어서, 프로모터 서열은 혐기조건하에서 코딩서열의 발현을 촉진시킬 수 있는 nirB 프로모터 또는 이의 유도체인 것을 특징으로 하는 DNA 구조.
  10. 전술한 어느 한항에 있어서, 제2단백질은 바이러스, 박테리아, 곰팡이, 이스트 또는 기생충에서 유도된 항원서열인 것을 특징으로 하는 DNA 구조.
  11. 제7항에 있어서, 항원서열은 쉬스토소마 만소니 또는 이의 에피토프 또는 사람 유두종 바이러스, 허피스 심플렉스 바이러스, FMDS 또는 SIDV 바이러스에서 유도한 항원서열을 구성되는 것을 특징으로 하는 DNA 구조.
  12. 주변환경의 변화에 상응하여 활성이 유도되는 프로모터 서열로 구성된 DNA 구조에서 프로모터 서열은 제1항원성 서열과 힌지부분과 그리고 이의 3' 단부에 인접하여 제2항원성 서열의 유도를 위한 하나이상의 제한 효소 부위를 인코드하는 DNA 서열에 작동식으로 연결되는 것을 특징으로 하는 DNA 구조.
  13. 제12항에 따른 힌지부분, 파상풍 독소단편 C 또는 이의 하나이상의 에피토프를 인코드하는 제1DNA서열에 연결된 프로모터로 구성된 것을 특징으로 하는 DNA 구조.
  14. 제13항에 있어서, 프로모터는 주변환경의 변화에 반응하여 유도되는 활성을 가지는 것을 특징으로 하는 DNA 구조.
  15. 제12항 내지 14항중 어느 한항에 있어서, 프로모터 서열은 혐기성 조건하에서 유도되는 활성인 것을 특징으로 하는 DNA 구조.
  16. 제15항에 있어서, 프로모터는 혐기상태에서 서열의 발현을 촉진시키는 능력이 있는 nirB 프로모터 또는 이의 일부분 또는 유도체인 것을 특징으로 하는 DNA 구조.
  17. 제1 내지 16항중 어느 한항에 따른 DNA 구조를 포함하고 박테리아에서 사용할 수 있는 복제가능한 발현 벡터.
  18. 제17항에 따른 박현벡터로 형질전환된 박테리아.
  19. 제18항에 있어서, 박테리아는 감쇠된 것을 특징으로 하는 박테리아.
  20. 제1항 내지 18항에 따른 DNA 구조로 감쇠 박테리아내로 형질 도입시키는 것으로 구성된 제19항에 따른 감쇠 박테리아 제조과정.
  21. 힌지부분에 의해 연결된 제1과 제2단백질로 구성된 융합단백질에 있어서, 융합단백질은 제17항에 따른 복제가능한 발현벡터에 의해 발현되는 것을 특징으로 하는 융합 단백질.
  22. 파상풍 독소 단편 C 또는 이의 하나이상의 에피토프와 제2이형 단백질이 연결된 것으로 구성된 것을 특징으로 하는 융합 단백질.
  23. 제22항에 있어서, 실제 순수형인 것을 특징으로 하는 융합 단백질.
  24. 제19항에 따른 감쇠박테리아 또는 제22 또는 23항에 따른 융합단백질 그리고 제약학적으로 수용가능한 담체로 구성된 것을 특징으로 하는 백신 조성물.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019950700271A 1992-07-31 1993-07-30 감쇠 박테리아에서 재조합 융합 단백질의 발현(Expression of recombinant fusion proteins in attenuated bacteria) KR950702635A (ko)

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GB9216317.9 1992-07-31
GB929216317A GB9216317D0 (en) 1992-07-31 1992-07-31 Vaccines
GB939306398A GB9306398D0 (en) 1993-03-26 1993-03-26 Vaccines
GB9306398.0 1993-03-26
PCT/GB1993/001617 WO1994003615A1 (en) 1992-07-31 1993-07-30 Expression of recombinant fusion proteins in attenuated bacteria

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JPH09500540A (ja) * 1993-07-30 1997-01-21 メデヴァ ホールディングス ビー．ヴイ． ワクチン組成物
GB9401787D0 (en) 1994-01-31 1994-03-23 Medeva Holdings Bv Vaccine compositions
GB2295394B (en) * 1994-01-31 1997-09-24 Medeva Holdings Bv DNA encoding a fusion protein comprising the C fragment of tetanus toxin
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US6680182B1 (en) 2004-01-20
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ATE174628T1 (de) 1999-01-15
FI950396A0 (fi) 1995-01-30
CA2141427C (en) 2008-07-22
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EP0652962B1 (en) 1998-12-16
EP0863211A1 (en) 1998-09-09
NO950348L (no) 1995-03-28
FI950396A (fi) 1995-01-30
AU4719393A (en) 1994-03-03
DE69322645D1 (de) 1999-01-28
JP3633933B2 (ja) 2005-03-30
JPH08503602A (ja) 1996-04-23
WO1994003615A1 (en) 1994-02-17
DK0652962T3 (da) 1999-08-23
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