KR890010198A - 효모에서 외래단백질을 제조하는 방법, 재조합 dna 및 형질전환체 - Google Patents

효모에서 외래단백질을 제조하는 방법, 재조합 dna 및 형질전환체 Download PDF

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KR890010198A
KR890010198A KR1019880005553A KR880005553A KR890010198A KR 890010198 A KR890010198 A KR 890010198A KR 1019880005553 A KR1019880005553 A KR 1019880005553A KR 880005553 A KR880005553 A KR 880005553A KR 890010198 A KR890010198 A KR 890010198A
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serum albumin
signal peptide
gene
foreign protein
ser
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KR1019880005553A
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KR970005250B1 (ko
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나오푸미 하야수께
유끼미쯔 나까가와
유따까 이시다
켄 오카바야시
코지 무라까미
기요시 쯔쭈이
가주오 이께가야
히도시 미나미노
사다오 우에다
하루히다 가와베
히로부미 아리무라
아쯔시 마사끼
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수야마 다다가즈
가부시끼가이샤 미도리쥬지
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/76—Albumins
    • C07K14/765—Serum albumin, e.g. HSA
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
    • C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence

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Abstract

내용 없음

Description

효모에서 외래단백질을 제조하는 방법, 재조합 DNA 및 형질전환체
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제1도는 GAL1, 10 프로모터를 함유하는 pGAL11로부터 pGAL12를 제조하는 과정을 나타낸다.
제2도는 Pho5 유전자 전부를 함유하는 pAP5 및 pUC9로부터, Pho5 터미네이터만을 함유하는 pPT1을 제조하는 과정을 나타낸다.

Claims (10)

  1. 외래단백질 유전자에 인접한 혈청알부민 시그널 펩티드 유전자, 혈청 알부민 시그널 펩티드 유전자에 대해 상부에 위치한 프로모터 및 외래 단백질 유전자에 대해 하부에 위치한 터미네이터를 함유하는 재조합 DNA로 형질전환된 효모에 의해 언급된 외래 단백질을 발현하고 분비시킴을 특징으로하여 외래 단백질을 제조하는 방법.
  2. 제1항에 있어서, 혈청알부민 시그널 펩티드가 인간에서 유래된 방법.
  3. 제1항에 있어서, 혈청알부민 시그널 펩티드 유전자가 하기 아미노산 서열로 발현되는 방법.
    Met Y Trp Val Tr Phe Ile Ser Leu Leu Phe Leu Phe X5X4X3X2X1
    상기 서열에서, Y는 Lys, Arg 또는 His를 나타내고 ; X5는 Ala, Pro 또는 Ser을 나타내며 ; X4는 Lys, Gly 또는 Ser을 나타내고 ; X3는 Ala, Val 또는 Cys를 나타내며 ; X2는 Tyr, Trp 또는 Ser을 나타내고 ; X1은 Ser, Ala 또는 Gly을 나타낸다.
  4. 하기 아미노산 서열을 암호화하는 혈청알부민 시그널 펩티드 유전자.
    Met Y Trp Val Thr Phe Ile Ser Leu Leu Phe Leu Phe X5X4X3X2X1
    상기 서열에서, Y는 Lys, Arg 또는 His를 나타내고 ; X5는 Ala, Pro 또는 Ser을 나타내며 ; X4는 Lys, Gly 또는 Ser을 나타내고 ; X3는 Ala, Val 또는 Cys를 나타내며 ; X2는 Tyr, Trp 또는 Ser을 나타내고 ; X1은 Ser, Ala 또는 Gly을 나타낸다.
  5. 외래 단백질을 암호화하는 DNA에 인접한 혈청알부민 시그널 펩티드를 암호화하는 DNA, 언급한 혈청알부민 시그널 펩티드 유전자에 대해 상부에 위치한 프로모터 및 언급한 외래 단백질 유전자에 대해 하부에 위치한 터미네이터를 함유하는, 효모를 형질전환시키기 위한 재조합 DNA.
  6. 외래 단백질을 암호화하는 DNA에 인접한 혈청알부민 시그널 펩티드를 암호화하는 DNA, 언급한 혈청알부민 시그널 펩티드 유전자에 대해 상부에 위치한 프로모터 및 언급한 외래 단배질 유전자에 대해 하부에 위치한 터미네이터를 함유하는 재조합 DNA에 의해 형질전환된 효모 균주.
  7. 제1항에 있어서, 언급한 혈청 알부민 시그널 펩티드의 아미노산 서열이 하기 그룹으로부터 선택되는 방법 :
    MetLysTrpValThrPheIleSerLeuLeuPheLeuPheSerSerAlaTyrSer, MetLysTrpValThrPheLeuLeuLeuLeuPheIleSerGlySerAlaPheSer, MetLysTrpValThrPheIleSerLeuLeuLeuLeuPheserSerAlaTurSer, MetLysTrpValThrPheIleSerLeuLeuPheLeuPheSerSerValTyrAla, MetLysTrpValThrPheIleSerLeuLeuPheLeuPheAlaLysValSerAla, MetLysTrpValThrPheIleSerLeuLeuPheLeuPheProGlyCysTrpAla, 및 MetLysTrpValThrPheIleSerLeuLeuPheLeuPheProGlyValTrpAla.
  8. 제1항에 있어서, 언급한 혈청 알부민 시그널 펩티드 유전자가 하기의 DNA 서열을 갖는 방법.
    ATGAAGTGGGTAACCTTTATTTCCCTTCTTTTTCTCTTTAGCTCGGCTTATTCC.
  9. 제1항에 있어서, 언급한 외래 단백질이 인간 혈청알부민, 인터페론-α, 인터페론-β,인터페론-, 유로키나제, 성장호르몬, 인슐린, 림포킨, h-ANP, 인자Ⅷ, CSFS 및 EPO로 구성된 그룹으로 부터 선택되는 방법.
  10. 제1항에 있어서, 외래 단백질 유전자가 혈청 알부민 시그널 펩티드 유전자의 바로 하부에 위치하는 완전한 인간 혈청 알부민 유전자인 방법.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019880005553A 1987-12-02 1988-05-13 효모에서 외래단백질을 제조하는 방법 및 재조합 dna KR970005250B1 (ko)

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JP30667487 1987-12-02
JP4560588 1988-02-26
JP63-45605 1988-02-26
JP62-306674 1988-02-26

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EP (1) EP0319641B1 (ko)
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CA (1) CA1326217C (ko)
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ES2059428T3 (es) 1994-11-16
KR970005250B1 (ko) 1997-04-14
JPH02167095A (ja) 1990-06-27
EP0319641B1 (en) 1993-09-22
DE3884356D1 (de) 1993-10-28
CA1326217C (en) 1994-01-18
US5503993A (en) 1996-04-02
DE3884356T2 (de) 1994-03-10
EP0319641A1 (en) 1989-06-14
JP2791418B2 (ja) 1998-08-27

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