KR950701348A - 재조합 사람 락토페린 생산방법 - Google Patents

재조합 사람 락토페린 생산방법

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Publication number
KR950701348A
KR950701348A KR1019940703776A KR19940703776A KR950701348A KR 950701348 A KR950701348 A KR 950701348A KR 1019940703776 A KR1019940703776 A KR 1019940703776A KR 19940703776 A KR19940703776 A KR 19940703776A KR 950701348 A KR950701348 A KR 950701348A
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South Korea
Prior art keywords
plasmid
eukaryotic cell
lactophrine
glucoamylase
amylase
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KR1019940703776A
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English (en)
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KR0184693B1 (ko
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오로라 엠. 코네릴
데니스 알. 헤돈
버트 더블유. 오'말리
그레고리 에스. 메이
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사무엘 에스 크로커
베이롤 칼리지 오브 메드신
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Publication of KR950701348A publication Critical patent/KR950701348A/ko
Priority to KR1019980707113A priority Critical patent/KR100221404B1/ko
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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
    • 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
    • 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
    • C07K14/79—Transferrins, e.g. lactoferrins, ovotransferrins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • Biophysics (AREA)
  • Biomedical Technology (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Mycology (AREA)
  • Toxicology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Peptides Or Proteins (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

본 발명은 신규한 플라스미드 형질전환된 진핵세포, 이 플라스미드와 진핵세포의 생성방법을 제공한다. 신규한 플라스미드는 사람 락토폐린 단백질의 cDNA를 포함한다. A. 오리자에서 사람 락토폐린 단백질의 생산방법이 제공된다.
따라서 본 발명은 재조합 사람 락토폐린 단백질의 생산을 위한 효과적이고 경제적인 수단을 제공한다.

Description

재조합 사람 락토페린 생산방법
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제1도는 아스피라질러스 오리자 발현 플라스미드 pAhlfg의 구조이다.
제3도는 형질전환체와 콘트롤 A07의 RNA분석을 설명한다.

Claims (21)

  1. 생물학적으로 활성이 있는 재조합 락토페린을 생산하는 방법에 있어서, 선택성 표식유전자, 프로모터, 전사종료서열과 링커(linker) 서열을 복합시키고; 플라스미드를 만들기 위해 서열을 클로닝시키고; 제한 효소로 플라스미드를 절단시키고; 제한효소 부위에 락토폐린 cDNA 코드를 삽입시키고; 그리고 락토폐린 cDNA를 발현시키는 플라스미드를 진핵세포에 형질도입시키는 단계로 구성된 것을 특징으로 하는 방법.
  2. 제1항에 있어서, 선택성 표식유전자는 pyr4, pyrG, andS, argB와 trpC에서 선택되는 것을 특징으로 하는 방법.
  3. 제1항에 있어서, 세포가 락토폐린을 발현시키는 것을 특징으로 하는 방법.
  4. 제2항의 방법에 따라 생성된 것을 특징으로 하는 락토폐린.
  5. 제1항에 있어서, 프로모터는 알코올 가수분해효소, argB, α-아밀라제, 글루코아밀라제와 benA에서 선택되는 것을 특징으로 하는 방법.
  6. 제1항에 있어서, 전사종료서열은 α-아밀라제, 글루코아밀라제, 알코올 가수분해효소 benA에서 선택되는 것을 특징으로 하는 방법.
  7. 제1항에 있어서, 링커서열은 α-아밀라제, 글루코아밀라제와 락토폐린에서 선택되는 것을 특징으로 하는 방법.
  8. 도면 6의 cDNA로 구성되고 진핵세포에서 cDNA를 발현시키는데 필요한 조절요소로 구성된 진핵세포에서 발현하기에 적합한 것을 특징으로 하는 플라스미드.
  9. pAhLFG플라스미드.
  10. 제8항에 따른 플라스미드를 포함하는 것을 특징으로 하는 진핵세포.
  11. 제10항에 있어서, 진핵세포는 곰팡이와 곤충세포에서 선택되는 것을 특징으로 하는 진핵세포.
  12. 제11항에 있어서, 곤충세포는 SF9인 것을 특징으로 하는 진핵세포.
  13. 제11항에 있어서, 곰팡이는 이스트인 것을 특징으로 하는 진핵세포.
  14. 제13항에 있어서, 이스트가 아스퍼질러스인 것을 특징으로 하는 진핵세포.
  15. 제14항에 있어서, 아스퍼질러스 균주는 A. 오리자(ortzae), A. 나이저(Niger), A. 니듈란(Nidulans0과 A. 오와모리(Awamori)에서 선택되는 것을 특징으로 하는 진핵세포.
  16. 재조합 플라스미드를 포함하는 형질전환체 진핵세포를 락토폐닌을 생산하는 공정에 있어서, 플라스미드는 락토폐린 단백질을 코드하는 폴리데옥시 리보뉴클레오티드를 가지는 플라스미드 벡터로 구성되고, 이는 적절한 염 양배지에서 락토폐린 단백질이 형성될때까지 배양하여 사람 락토폐린을 분리하는 단계로 구성된 것을 특징으로 하는 공정.
  17. (1)유전자 발현에서 조절역할을 하는 유전자 요소; (2)사람 락토폐린에 대한 cDNA코드; 그리고(3)적절한 전사와 해독개시와 종료서열로된 전사유닛을 가지는 것을 특징으로 하는 재조합 발현벡터.
  18. 제17항에 있어서, 유전자 요소가 프로모터인 것을 특징으로 하는 재조합 발현벡터.
  19. 제18항에 있어서, 프로모터는 알코올 분해효소, argB, α-아밀라제, 글루코아밀라제와 benA에서 선택되는 것을 특징으로 하는 재조합 발현벡터.
  20. 제17항에 있어서, 전사종료서열을 알코올 분해효소, argB, α-아밀라제, 글루코아밀라제와 benA에서 선택되는 것을 특징으로 하는 재조합 발현벡터.
  21. 제16항에 따라 생성된 것을 특징으로 하는 단백질.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019940703776A 1992-04-24 1993-04-16 재조합 사람 락토페린 생산방법 KR0184693B1 (ko)

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US87330492A 1992-04-24 1992-04-24
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BR (1) BR9306289A (ko)
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NZ (1) NZ252844A (ko)
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JP2818056B2 (ja) * 1990-09-07 1998-10-30 森永乳業株式会社 抗菌性ペプチドおよび抗菌剤

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CN1047204C (zh) 1999-12-08
ATE177473T1 (de) 1999-03-15
CN1161382C (zh) 2004-08-11
CA2134094A1 (en) 1993-11-11
BR9306289A (pt) 1998-06-30
CA2134094C (en) 2004-01-06
IL105400A (en) 2003-03-12
DK0644899T3 (da) 1999-09-27
EP0644899B1 (en) 1999-03-10
JPH08504561A (ja) 1996-05-21
AU681583B2 (en) 1997-09-04
DE69323879T2 (de) 1999-11-04
US5571896A (en) 1996-11-05
IL105400A0 (en) 1993-08-18
CN1250055A (zh) 2000-04-12
EP0644899A1 (en) 1995-03-29
KR100221404B1 (en) 1999-10-01
ZA932568B (en) 1993-11-12
CN1079780A (zh) 1993-12-22
HK1027360A1 (en) 2001-01-12
SG47605A1 (en) 1998-04-17
GR3029942T3 (en) 1999-07-30
WO1993022348A1 (en) 1993-11-11
ES2132232T3 (es) 1999-08-16
DE69323879D1 (de) 1999-04-15
AU4289293A (en) 1993-11-29
EP0644899A4 (en) 1996-06-26
JP2824332B2 (ja) 1998-11-11
KR0184693B1 (ko) 1999-04-01

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