KR950005990A - Expression of human interleukin-4 in yeast - Google Patents

Expression of human interleukin-4 in yeast Download PDF

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Publication number
KR950005990A
KR950005990A KR1019930015350A KR930015350A KR950005990A KR 950005990 A KR950005990 A KR 950005990A KR 1019930015350 A KR1019930015350 A KR 1019930015350A KR 930015350 A KR930015350 A KR 930015350A KR 950005990 A KR950005990 A KR 950005990A
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South Korea
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expression vector
human interleukin
yeast
gene
interleukin
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KR1019930015350A
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Korean (ko)
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KR100274183B1 (en
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조중명
김성택
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최근선
주식회사 럭 키
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    • 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]
    • C07K14/5406—IL-4
    • 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
    • C07K2319/00—Fusion polypeptide
    • C07K2319/50—Fusion polypeptide containing protease site

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Biophysics (AREA)
  • Wood Science & Technology (AREA)
  • Molecular Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biochemistry (AREA)
  • Mycology (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Toxicology (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

본 발명은 유전공학적인 방법에 의해 제조된 인간 인터류킨-4유전자, 그를 포함하는 발현 벡터, 상기 발현벡터로 형질전환된 미생물 및 상기 미생물을 이용한 인터류킨-4의 생산 방법에 관한 것이다. 보다 상세하게는, IL-4 단백질의 아미노산 서열을 그대로 유지하면서 유전자의 대량 발현을 가능케 하는 코돈을 사용하여 설계된 인간 인터류킨-4유전자, 상기 유전자를 유비퀴틴 유전자와 결합된 상태로 포함하여 그의 발현시 인간 인터류킨-4와 유비퀴틴의 융합 단백질을 생산하는 발현 벡터, 상기 발현 벡터로 형질 전화된 미생물 및 상기 미생물을 배양함으로써 인간 인터류킨-4를 대량으로 생산하는 방법에 관한 것이다.The present invention relates to a human interleukin-4 gene produced by a genetic engineering method, an expression vector comprising the same, a microorganism transformed with the expression vector, and a method for producing interleukin-4 using the microorganism. More specifically, the human interleukin-4 gene, which is designed using codons capable of mass expression of genes while maintaining the amino acid sequence of the IL-4 protein, comprising the gene in combination with the ubiquitin gene, and human An expression vector for producing a fusion protein of interleukin-4 and ubiquitin, a microorganism transformed with the expression vector, and a method for producing a large amount of human interleukin-4 by culturing the microorganism.

Description

인간 인터류킨-4(human interleukin-4)의 효모에서 발현Expression in the yeast of human interleukin-4

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

제1도는 5'-말단에 Sacll 제한 효소 부위, 3'-말단에 Sall 제한 효소 부위를 갖는 IL-4유전자의 염기 서열및 아미노산 서열을 나타낸 것이고,1 shows the nucleotide sequence and amino acid sequence of the IL-4 gene having a Sacll restriction enzyme site at the 5'-end and a Sall restriction enzyme site at the 3'-end,

제2도는 전체 IL-4유전자의 합성올리고머 1,2,3,4,5,6,7,8,9 및 10의 위치 및 그연결 순서를 나타낸 것이고,Figure 2 shows the positions of the synthetic oligomers 1,2,3,4,5,6,7,8,9 and 10 of the whole IL-4 gene and their sequence of connection,

제3도는 IL-4 유전자의 M13 백터로의 클로닝 및 효모발현 벡터로의 클로닝을 도식화한 것이고,3 shows the cloning of IL-4 gene into M13 vector and cloning into yeast expression vector,

제4도는 본 발명의 IL-4 발현 벡터(pYLBC-A/G-UB-IL-4)를 함유하는 효모의 농축 배양액을 SDS 폴리아크릴아미드 젤 전기 영동한 결과를 나타낸 것이다.Figure 4 shows the results of SDS polyacrylamide gel electrophoresis of the concentrated culture of yeast containing the IL-4 expression vector (pYLBC-A / G-UB-IL-4) of the present invention.

Claims (6)

5' 말단에 Sacll 제한 효소 인지부위를 포함하고 3'-말단에 Sall 제한 효소 인지부위를 포함하는, 인간 인터류킨-4을 코딩하는 하기의 뉴클레오티드 서열을 포함하는 DNA.A DNA comprising the following nucleotide sequence encoding human interleukin-4, comprising a Sacll restriction enzyme recognition site at the 5 'end and a Sall restriction enzyme recognition site at the 3'-end. 제1항의 DNA를 포함하는 발현벡터.An expression vector comprising the DNA of claim 1. 제2항에 있어서, pYLBC-A/G-UB-IL-4인 발현벡터.The expression vector according to claim 2, which is pYLBC-A / G-UB-IL-4. 제2항의 발현벡터로 형질전환된 효모.Yeast transformed with the expression vector of claim 2. 제4항에 있어서, 발현벡터 pYLBC-A/G-UB-IL-4로 형질전환된 효모(KCTC 0069BP).The yeast (KCTC 0069BP) transformed with the expression vector pYLBC-A / G-UB-IL-4. 제4항 또는 제5항의 효모를 배양함을 포함하는, 인간 인터류킨-4의 생산 방법.A method for producing human interleukin-4, comprising culturing the yeast of claim 4 or 5. ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.※ Note: The disclosure is based on the initial application.
KR1019930015350A 1993-08-07 1993-08-07 Expression of human interleukin-4 in yeast KR100274183B1 (en)

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Application Number Priority Date Filing Date Title
KR1019930015350A KR100274183B1 (en) 1993-08-07 1993-08-07 Expression of human interleukin-4 in yeast

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Application Number Priority Date Filing Date Title
KR1019930015350A KR100274183B1 (en) 1993-08-07 1993-08-07 Expression of human interleukin-4 in yeast

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KR950005990A true KR950005990A (en) 1995-03-20
KR100274183B1 KR100274183B1 (en) 2000-12-15

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100443570B1 (en) * 2001-07-31 2004-08-09 크레아젠 주식회사 The preparation method of recombinant human Interleukin-4

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100443570B1 (en) * 2001-07-31 2004-08-09 크레아젠 주식회사 The preparation method of recombinant human Interleukin-4

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