KR950005989A - Expression of human interleukin-3 in yeast - Google Patents

Expression of human interleukin-3 in yeast Download PDF

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Publication number
KR950005989A
KR950005989A KR1019930015349A KR930015349A KR950005989A KR 950005989 A KR950005989 A KR 950005989A KR 1019930015349 A KR1019930015349 A KR 1019930015349A KR 930015349 A KR930015349 A KR 930015349A KR 950005989 A KR950005989 A KR 950005989A
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KR
South Korea
Prior art keywords
expression vector
yeast
human interleukin
gene
expression
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KR1019930015349A
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Korean (ko)
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KR100274182B1 (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]
    • C07K14/5403—IL-3
    • 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

Abstract

본 발명은 유전공학적인 방법에 의해 제조된 인간 인터류킨-3 유전자, 그를 포함하는 발현 벡터, 상기 발현벡터로 형질전환된 미생물 및 상기 미생물을 이용한 인터류킨-3의 생산 방법에 관한 것이다. 보다 상세하게는 IL-3 단백질의 아미노산 서열은 그대로 유지하면서 유전자의 대량 발현을 가능케 하는 코돈을 사용하여 설계된 인간 인터류킨-3 유전자, 상기 유전자를 포함하는 발현 벡터, 상기 발현 벡터로 형질 전환된 효모 및 상기 효모를 배양함으로써 인간 인터류킨-3을 대량으로 생산하는 방법에 관한 것이다.The present invention relates to a human interleukin-3 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-3 using the microorganism. More specifically, the human interleukin-3 gene, which is designed using codons that allow mass expression of the gene while maintaining the amino acid sequence of the IL-3 protein, an expression vector comprising the gene, a yeast transformed with the expression vector, and It relates to a method for mass production of human interleukin-3 by culturing the yeast.

Description

인간 인터류킨-3(human interleukin-3)의 효모에서의 발현Expression of human interleukin-3 in yeast

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

제1도는 IL-3 유전자의 5'-말단에 Xbal 제한효소 인지 부위를, 3'-말단에 종료코돈과 제한효소 Sall 인지부위를 갖도록 설계된 IL-3 유전자의 뉴클레오티드서열 및 아미노산 서열을 나타낸 것이고,Figure 1 shows the nucleotide sequence and amino acid sequence of the IL-3 gene designed to have the Xbal restriction enzyme recognition site at the 5'-end of the IL-3 gene, the end codon and the restriction enzyme Sall recognition site at the 3'-end,

제2도는 전체 IL-3 유전자의 합성 올리고머 1,2,3,4,5,6,7,8,9 및 10의 위치 및 그 접합 순서를 나타낸 것이고,Figure 2 shows the positions of the synthetic oligomers 1,2,3,4,5,6,7,8,9 and 10 and the conjugation sequence of the whole IL-3 gene,

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

제4도는 본 발명의 IL-3 발현 벡터(pYLBC-A/G-αF-IL-3)를 함유하는 효모의 농축 배양액을 SDS 폴리아크릴아미드 젤 전기영동한 결과를 나타낸 것이다.4 shows the results of SDS polyacrylamide gel electrophoresis of a concentrated culture solution of yeast containing the IL-3 expression vector (pYLBC-A / G-αF-IL-3) of the present invention.

Claims (6)

5' 말단에 Xbal 제한 효소 인지부위를 포함하고 3'-말단에 Sall 제한 효소 인지부위를 포함하는, 인간 인터류킨-3을 코딩하는 하기의 뉴클레오티드 서열을 포함하는 DNA.A DNA comprising the following nucleotide sequence encoding human interleukin-3 comprising a Xbal 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-αF-IL-3인 발현벡터.The expression vector according to claim 2, which is pYLBC-A / G-αF-IL-3. 제2항의 발현벡터로 형질전화된 효모.Yeast transformed with the expression vector of claim 2. 제4항에 있어서, 발현벡터 pYLBC-A/G-αF-IL-3으로 형질전환된 효모(KCTC 0071BP).The yeast transformed with the expression vector pYLBC-A / G-αF-IL-3 (KCTC 0071BP). 제4항 또는 제5항의 효모를 배양함을 포함하는, 인간 인터류킨-3의 생산방법.A method for producing human interleukin-3, comprising culturing the yeast of claim 4. ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.※ Note: The disclosure is based on the initial application.
KR1019930015349A 1993-08-07 1993-08-07 Expression of human interleukin-3 in yeast KR100274182B1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
KR1019930015349A KR100274182B1 (en) 1993-08-07 1993-08-07 Expression of human interleukin-3 in yeast

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
KR1019930015349A KR100274182B1 (en) 1993-08-07 1993-08-07 Expression of human interleukin-3 in yeast

Publications (2)

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KR950005989A true KR950005989A (en) 1995-03-20
KR100274182B1 KR100274182B1 (en) 2000-12-15

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KR1019930015349A KR100274182B1 (en) 1993-08-07 1993-08-07 Expression of human interleukin-3 in yeast

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100819314B1 (en) * 2001-03-28 2008-04-02 주식회사 케이티 The analysis methods for call of Public Switch Telephone Network

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100819314B1 (en) * 2001-03-28 2008-04-02 주식회사 케이티 The analysis methods for call of Public Switch Telephone Network

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