KR960017850A - New yeast expression vector containing modified promoter and 1eu2-d gene - Google Patents

New yeast expression vector containing modified promoter and 1eu2-d gene Download PDF

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KR960017850A
KR960017850A KR1019940029992A KR19940029992A KR960017850A KR 960017850 A KR960017850 A KR 960017850A KR 1019940029992 A KR1019940029992 A KR 1019940029992A KR 19940029992 A KR19940029992 A KR 19940029992A KR 960017850 A KR960017850 A KR 960017850A
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expression vector
pyhil
pyh
dh5α
gene
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KR1019940029992A
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KR0141312B1 (en
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유향숙
정경숙
최원자
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김은영
한국과학기술연구원
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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
    • 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
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • 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
    • C07K14/52—Cytokines; Lymphokines; Interferons
    • C07K14/54—Interleukins [IL]
    • C07K14/5412—IL-6
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • 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
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16051—Methods of production or purification of viral material

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Abstract

본 발명은 프로모터에 연결된 단백질 유전자의 번역 개시 코돈으로부터 세번째 앞의 염기가 아데닌으로 치환된 변형된 프로모터와 효모의 루이신 합성에 관여하는 LEU2 유전자의 프로머터가 일부 제거된 leu2-d 유전자를 포함하는, 외래 유전자의 효모에서 발현율을 높일 수 있는 신규 발현 벡터에 관한 것이다.The present invention includes a modified promoter in which a third preceding base is substituted with adenine from a translation initiation codon of a protein gene linked to a promoter, and a leu2-d gene in which a part of the promoter of the LEU2 gene involved in leucine synthesis of yeast is partially removed. It relates to a novel expression vector that can increase the expression rate in yeast of foreign genes.

Description

변형된 프로모터와 leu2-d 유전자를 포함하는 신규 효모 발현 벡터New yeast expression vector containing modified promoter and leu2-d gene

본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음As this is a public information case, the full text was not included.

Claims (11)

외매 목적 단백질을 코드하는 유전자 및 상기 유전자의 번역 개시 코돈으로부터 상류쪽으로 3번재의 염기가 아데닌이 되도록 변형시킨 프로코터를 포함하는, 효모내에서 외래 목적 단백질을 대량으로 발현시키기 위한 발현 벡터.An expression vector for expressing a large amount of a foreign target protein in yeast, comprising a gene encoding a foreign protein of interest and a coater modified so that the third base is adenine upstream from the translation start codon of the gene. 제1항에 있어서, 상기 발현 벡터의 상기 변형된 프로모터가 3′ -말단의 염기 서열이 5′ -CAATAGG-3′인 변형된 PGK 프로모터 도는 변형된 칼라틴 프로모터인 발현 벡터.The expression vector of claim 1, wherein said modified promoter of said expression vector is a modified PGK promoter or modified colorin promoter having a 3′-terminal nucleotide sequence of 5′-CAATAGG-3 ′. 제1항에 있어서, 효모세포내에서 플라스미드의 수를 증가시킬 수 있는 leu2-d 유전자를 추가로 포함하는 발현 벡터.The expression vector of claim 1, further comprising a leu2-d gene capable of increasing the number of plasmids in yeast cells. 제2항 또는 제3항에 있어서, 상기 목적 단백질이 인간 면역 결핍 바이러스(HIV-I)의 표면 단백질 P17 또는 인터루킨-6 인 발현 벡터.The expression vector according to claim 2 or 3, wherein the target protein is the surface protein P17 or interleukin-6 of human immunodeficiency virus (HIV-I). 제4항에 있어서, 플라스미드 pYH-10ZM, pYH-10Zd, pYH-FZM, pYH-FZd, pYHIL-10ZM, pYHIL-10Zd, pYHIL-FZM 또는 pYHIL-FZd인 발현 벡터.The expression vector of claim 4, which is plasmid pYH-10ZM, pYH-10Zd, pYH-FZM, pYH-FZd, pYHIL-10ZM, pYHIL-10Zd, pYHIL-FZM or pYHIL-FZd. 제1항 내지 제5항 중 어느 한 항의 발??현 벡터로 형질 전환된 미생물.A microorganism transformed with the expression vector of any one of claims 1 to 5. 제6항에 있어서, 제5항의 플라스미드로 형질전환된 대장균 DH5α/pYH-10ZM, DH5α/pYH-10Zd(KCTC 8629P), DH5α/pYH-FZd (KCTC 8628P), DH5α/pYHIL-10ZM, DH5α/pYHIL-10Zd, DH5α/pYHIL-FZM 또는 DH5α/pYHIL-FZd.The E. coli DH5α / pYH-10ZM, DH5α / pYH-10Zd (KCTC 8629P), DH5α / pYH-FZd (KCTC 8628P), DH5α / pYHIL-10ZM, DH5α / pYHIL transformed with the plasmid of claim 5 -10Zd, DH5α / pYHIL-FZM or DH5α / pYHIL-FZd. 제6항에 있어서, 형질전환된 효모세포.The yeast cell of claim 6, which is transformed. 제8항에 있어서, 제5항의 발현 벡터로 형질전환된 효모세포 20B-12 또는 L3262.The yeast cell 20B-12 or L3262 of Claim 8 transformed with the expression vector of Claim 5. 제8항의 형질 전환된 효모 세포를 배양하고 배양된 세포 추출물로부터의 외래 단백질을 분리함을 포함하는, 상기 외래 단백질의 제조 방법.A method for producing the foreign protein, comprising culturing the transformed yeast cell of claim 8 and separating the foreign protein from the cultured cell extract. 제10항에 있어서, 제9항의 효모 세포를 배양하고 배양된 세포 추출물로부터 HIV-I의 표면 단백질 P17 또는 인터루킴-6을 분리함을 포함하는, HIV-I의 표면 단백질 P17 또는 인터루킨-6의 제조 방법.The surface protein P17 or interleukin-6 of HIV-I according to claim 10, comprising culturing the yeast cells of claim 9 and isolating surface protein P17 or interleukin-6 of HIV-I from the cultured cell extract. Manufacturing method. ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.※ Note: The disclosure is based on the initial application.
KR1019940029992A 1994-11-16 1994-11-16 Novel expression vectors for yeast comprising modified promotor and leu2-d gene KR0141312B1 (en)

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