KR830003574A - 변칙합성 유전자의 미생물을 통한 발현 - Google Patents
변칙합성 유전자의 미생물을 통한 발현 Download PDFInfo
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- KR830003574A KR830003574A KR1019800002658A KR800002658A KR830003574A KR 830003574 A KR830003574 A KR 830003574A KR 1019800002658 A KR1019800002658 A KR 1019800002658A KR 800002658 A KR800002658 A KR 800002658A KR 830003574 A KR830003574 A KR 830003574A
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- 238000003786 synthesis reaction Methods 0.000 title claims description 4
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Classifications
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- 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
- C12N15/66—General methods for inserting a gene into a vector to form a recombinant vector using cleavage and ligation; Use of non-functional linkers or adaptors, e.g. linkers containing the sequence for a restriction endonuclease
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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
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- C—CHEMISTRY; METALLURGY
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- 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
- C07K14/575—Hormones
- C07K14/61—Growth hormones [GH] (Somatotropin)
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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
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- 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
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- 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
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- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/62—DNA sequences coding for fusion proteins
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- 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
- C12N15/70—Vectors or expression systems specially adapted for E. coli
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- 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
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
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- 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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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/50—Fusion polypeptide containing protease site
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- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
- C07K2319/74—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor
- C07K2319/75—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor containing a fusion for activation of a cell surface receptor, e.g. thrombopoeitin, NPY and other peptide hormones
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/185—Escherichia
- C12R2001/19—Escherichia coli
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S930/00—Peptide or protein sequence
- Y10S930/01—Peptide or protein sequence
- Y10S930/12—Growth hormone, growth factor other than t-cell or b-cell growth factor, and growth hormone releasing factor; related peptides
Abstract
내용 없음
Description
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
도면 1 : 인간의 성장호르몬의 처음 24개의 아미노산에 대한 유전자와 출발신호인 ATG 그리고 클로닝시에 필요한 링커를 제조하는 데 대한 합성도이다. 상부가닥(암호용)과 하부 가닥(보완용) 중에 있는 화살표는 유전자 단편을 제조키 위해 결합된 올리고뉴클레오타이드를 나타내고 있다.
도면 2 : 도면 1의 유존자단편을 형성키 위한 "U"와 "L" 올리고뉴클레오타이드의 결합체와 이것을 플라스미드클로닝 담체로 삽입하는 것이 나타나 있다.
도면 3 : 뇌하수체 mRNA 복사의 Hae Ⅲ 제한 효소에 대한 DNA 순서(암호 가닥만)와 이들이 암호하는 인간성장 호르몬의 아미노산이 나타나 있다. 비번역 mRNA에 대한 DNA("중지"에 연속)에서 처럼 주요한 제한부위가 나타나 있다.
도면 4 : 합성으로 제조하지 않은 인간의 성장호르몬의 아미노산에 대한 유전자 단편에 대한 클로닝 담체 제조와 인간 뇌하수체원에서 분리된 mRNA로 부터 역복사법에 의해 보완 DNa 로서의 유전자단편의 제조에 관한 것이다.
도면 5 : 박테리아에서 도면 2와 3의 플라스미드로 시작하는 인간성장호르몬을 발현할 수 있는 플라스미드의 제조에 대해 나타나 있다.
Claims (2)
- 아미노산 순서가 알려진 폴리펩타이이드에 대한 유전자 암호를 클로닝담체에 삽입하고 발현 푸로모터의 조절하에 미생물 계에서 발현시켜서 목적하는 폴리펩타이드를 다음과 같은 구체적인 방법을 통하여 제조하는 방법.a) mRNA를 역복사시켜서 적어도 상기 폴리펩타이드의 암호순서의 상당부분을 이루며, 목적폴리펩타이드보다는 발현산물에 대한 최초유전자의 단편을 얻는다.b) 만일에 최초단편에 상기 폴리펩타이드에 함유하는 이외의 아미노산에 대한 코돈이 들어있으면 적어도 상기암호순서의 상당부분을 함유하도록 하면서 이 불필요한 코돈을 어느 정도 제거하여 아직 상기 폴리펩타이드이외의 발현산물에 대한 단편을 얻는다.((a) 단계의 산물이나 필요할 때는 (b) 단계의 산물인 유전자 단편중에는 상기 폴리펩타이드의 모든 아미노산보다 적은 암호유전자가 함유될 수도 있다.)(c) 유기합성법에 의하여 상기 폴리펩타이드의 나머지 아미노산에 대한 유전자단편을 얻는데 이중에는 적어도 한단편에는 폴리펩타이드의 아미노만단 부위를 암호하는 유전자가 함유되어 있다.d) 상기 c)단계에서 유기합성법으로 얻은 유전자와 a)나 b)단계에 얻은 유전자를 적절히 해독할 수 있는 순서로 조합하고 발현 푸로모터의 조절하에 있도록 복사능이 있는 클로닝 담체에 장치하며, 이렇게하여 상기 폴리펩타이드의 아미노산 순서를 전부 발현시킬 수 있는 클로닝담체를 얻는다.
- 다음의 방법을 사용하여 인간의 성장호르몬을 제조 방법 :a) 통기와 진탕을 시키게 되어 있는 발효조 내의 수성의 영양분 함유발효액에다 인간의 성장호르몬을 발현시킬 수 있으며 복사능이 있는 플라스미드를 함유하는 박테리아 균주를 가하여b) 왕성한 성장을 유지시키기 위해 필요하면 영양분을 더 제공하고 통기와 진탕을 하면서 군주를 증식시켜서c) 결과적으로 얻어지는 미생물 세포들을 발효액으로 부터 분리하여d) 미생물 세포를 용균시켜e) 세포잔해를 상등액으로부터 분리하여f) 상등액중에 들어 있는 인간의 성장호르몬을 분리한다.※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1019860009542A KR870000701B1 (ko) | 1979-07-05 | 1986-11-12 | 인체 성장 호르몬의 제조방법 |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US55126 | 1979-07-05 | ||
US06/055,126 US4342832A (en) | 1979-07-05 | 1979-07-05 | Method of constructing a replicable cloning vehicle having quasi-synthetic genes |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
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KR1019860009542A Division KR870000701B1 (ko) | 1979-07-05 | 1986-11-12 | 인체 성장 호르몬의 제조방법 |
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Publication Number | Publication Date |
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KR830003574A true KR830003574A (ko) | 1983-06-21 |
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Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
KR1019800002658A KR830003574A (ko) | 1979-07-05 | 1980-07-04 | 변칙합성 유전자의 미생물을 통한 발현 |
KR1019860009542A KR870000701B1 (ko) | 1979-07-05 | 1986-11-12 | 인체 성장 호르몬의 제조방법 |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
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KR1019860009542A KR870000701B1 (ko) | 1979-07-05 | 1986-11-12 | 인체 성장 호르몬의 제조방법 |
Country Status (42)
Country | Link |
---|---|
US (7) | US4342832A (ko) |
EP (1) | EP0022242B1 (ko) |
JP (4) | JPH0612996B2 (ko) |
KR (2) | KR830003574A (ko) |
AR (1) | AR244341A1 (ko) |
AT (1) | ATE82324T1 (ko) |
AU (2) | AU533697B2 (ko) |
BE (1) | BE884012A (ko) |
BG (1) | BG41135A3 (ko) |
BR (1) | BR8008736A (ko) |
CA (2) | CA1164375A (ko) |
CH (1) | CH661939A5 (ko) |
CS (3) | CS250652B2 (ko) |
DD (3) | DD210071A5 (ko) |
DE (3) | DE3050722C2 (ko) |
DK (2) | DK173503B1 (ko) |
EG (1) | EG14819A (ko) |
ES (2) | ES493149A0 (ko) |
FI (2) | FI802030A (ko) |
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