KR960031601A - Vectors for Expression of Recombinant Human Antitrypsin-α-1 Protein from Yeast - Google Patents

Vectors for Expression of Recombinant Human Antitrypsin-α-1 Protein from Yeast Download PDF

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KR960031601A
KR960031601A KR1019950003140A KR19950003140A KR960031601A KR 960031601 A KR960031601 A KR 960031601A KR 1019950003140 A KR1019950003140 A KR 1019950003140A KR 19950003140 A KR19950003140 A KR 19950003140A KR 960031601 A KR960031601 A KR 960031601A
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South Korea
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vector
protein
human antitrypsin
expression
antitrypsin
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KR1019950003140A
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KR0145948B1 (en
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정봉현
남수완
박영훈
유명희
강현아
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김은영
한국과학기술연구원
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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
    • 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
    • C07K14/81—Protease inhibitors
    • C07K14/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
    • C07K14/811—Serine protease (E.C. 3.4.21) inhibitors
    • C07K14/8121—Serpins
    • C07K14/8125—Alpha-1-antitrypsin
    • 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
    • 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
    • C12N2800/00—Nucleic acids vectors
    • C12N2800/10—Plasmid DNA
    • C12N2800/102—Plasmid DNA for yeast

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  • Proteomics, Peptides & Aminoacids (AREA)
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Abstract

효모로부터 재조합 인간 안티트립신-α-1 단백질을 세로의 배지로 효율적으로 분비시키는 발현 분비 벡터에 관한 것으로, 이누리나제 분비 시그날 서열을 코딩하는 폴리뉴클레오티드와 연결된 인간 안티트립신-α-1 구조 유전자를 포함하는 인간 안티트립신-α-1의 발현 분비 벡터, 상기 발현 분비 벡터로 형질 전환된 재조합 효모 세포; 및 상기 재조합 효모 세포를 배양하여 인간 안티트립신-α-1 단백질을 생산하는 방법을 제공한다.An expression secretion vector that efficiently secretes recombinant human antitrypsin-α-1 protein from yeast into a longitudinal medium, wherein the human antitrypsin-α-1 structural gene is linked to a polynucleotide encoding an inulinase secretion signal sequence. Expression secretion vector of human antitrypsin-α-1, comprising recombinant yeast cells transformed with the expression secretion vector; And culturing the recombinant yeast cells to produce human antitrypsin-α-1 protein.

Description

효모로부터 재조합 인간 안티트립신-α-1 단백질을 발현 분비시키기 위한 벡터Vectors for Expression of Recombinant Human Antitrypsin-α-1 Protein from Yeast

본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음Since this is an open matter, no full text was included.

제1도는 본 발명에 따른 발현 분비 벡터 pYInu-AT의 제작과정을 나타낸 것이고,Figure 1 shows the manufacturing process of the expression secretion vector pYInu-AT according to the present invention,

제2도는 발현 분비 벡터 pYInu-AT 중의 이누리나제 분비 시그날 서열의 올리고뉴클레오티드의 구성도(A), 및 이누리나제 시그날 서열과 안티트립신-α-1의 연결부분의 뉴클레오티드 서열 및 아미노산 서열(B)을 나타낸 것이고,Figure 2 is a block diagram of the oligonucleotide of the inulinase secretion signal sequence in the expression secretion vector pYInu-AT (A), and the nucleotide sequence and amino acid sequence (B) of the linking portion of the inulinase signal sequence and antitrypsin-α-1 ),

제3도는 발현 분비 벡터 pYInu-AT를 함유하는 재조합 효모의 유가배양 상등액을 웨스턴 블롯팅(Western blotting)한 결과를 나타낸 것이다.FIG. 3 shows the result of Western blotting of the cultured supernatant of recombinant yeast containing the expression secreting vector pYInu-AT.

Claims (8)

이누리나제 분비 시그날 서열을 코딩하는 폴리뉴클레오티드와 연결된 인간 안티트립신-α-1 구조 유전자를 포함하는 인간 안티트립신-α-1 발현 분비 벡터.A human antitrypsin-α-1 expression secretion vector comprising a human antitrypsin-α-1 structural gene linked to a polynucleotide encoding an inulinase secretion signal sequence. 제1항에 있어서, 프로모터로서 갈락토스 유도성 GAL10 프로모터를 사용하는 벡터.The vector of claim 1, wherein a galactose inducible GAL10 promoter is used as a promoter. 제1항에 있어서, 상기 폴리뉴클레오티드가 클루이베로마이세스 마르시아누스(Kluyveromyces marxianus)의 이누리나제 분비 시그날 펩티드를 코팅하는 폴리뉴클레오티드인 벡터.The vector of claim 1, wherein the polynucleotide is a polynucleotide that coats the Inurinase secretion signal peptide of Kluyveromyces marxianus. 제1항에 있어서, 전사 종결 서열로서 GAL7 유전자 유래의 전사종결 서열을 사용하는 벡터.The vector according to claim 1, wherein the transcription termination sequence derived from the GAL7 gene is used as the transcription termination sequence. 제1항에 있어서, 플라스미스 pYInu-AT(KCTC 0132 BP).The plasmid pYInu-AT (KCTC 0132 BP) according to claim 1. 제1항 내지 제5항의 어느 한 항의 발현 분비 벡터로 형질전환된 재조합 효모.Recombinant yeast transformed with the expression secretion vector of any one of claims 1 to 5. 제6항에 있어서, 사카로마이세스 세레비지애 2805/pYInu-AT(KCTC 0132 BP).The method of claim 6, wherein Saccharomyces cerevisiae 2805 / pYInu-AT (KCTC 0132 BP). 제6항의 재조합 효모 세포를 배양 배지 상에서 배양한 후, 상기 배양 배지로부터 인간 안티트립신-α-1 단백질을 회수함을 포함하는, 인간 안티트립신-α-1 단백질의 생산 방법.A method for producing human antitrypsin-α-1 protein, comprising culturing the recombinant yeast cell of claim 6 on a culture medium, and then recovering human antitrypsin-α-1 protein from the culture medium. ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.※ Note: The disclosure is based on the initial application.
KR1019950003140A 1995-02-18 1995-02-18 VECTOR FOR EXPRESSION AND SECRETION OF RECOMBINANT HUMAN ANTITRYPSIN-Ñß-1 PROTEIN FROM YEAST KR0145948B1 (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100672745B1 (en) * 2005-04-20 2007-01-24 재단법인 목암생명공학연구소 Methods for enhancing a secretion efficiency of recombinant foreign protein in yeast expression system

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Publication number Priority date Publication date Assignee Title
KR102268749B1 (en) * 2020-05-06 2021-06-23 전북대학교산학협력단 Recombinant Yeast containing histamine-binding proteins and yeast surface expression protein

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100672745B1 (en) * 2005-04-20 2007-01-24 재단법인 목암생명공학연구소 Methods for enhancing a secretion efficiency of recombinant foreign protein in yeast expression system

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