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1. WO2013154312 - METHOD FOR PRODUCING A TOXIC PROTEIN BY USING LACI PROTEIN MUTANT

Publication Number WO/2013/154312
Publication Date 17.10.2013
International Application No. PCT/KR2013/002919
International Filing Date 08.04.2013
IPC
C07K 14/245 2006.01
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
KPEPTIDES
14Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
195from bacteria
24from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
245Escherichia (G)
C12N 15/63 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
C12N 15/70 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
70Vectors or expression systems specially adapted for E. coli
CPC
C07K 14/245
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
KPEPTIDES
14Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
195from bacteria
24from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
245Escherichia (G)
C12N 15/72
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
70Vectors or expression systems specially adapted for E. coli
72Expression systems using regulatory sequences derived from the lac-operon
C12P 21/02
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
21Preparation of peptides or proteins
02having a known sequence of two or more amino acids, e.g. glutathione
Applicants
  • 한국생명공학연구원 KOREA RESEARCH INSTITUTE OF BIOSCIENCE AND BIOTECHNOLOGY [KR]/[KR]
Inventors
  • 김지현 KIM, Jihyun F.
  • 윤성호 YOON, Sung Ho
  • 김성근 KIM, Seong Keun
  • 이대희 LEE, Dae-Hee
  • 권순경 KWON, Soon-Kyeong
  • 오태광 OH, Tae Kwang
  • 김오철 KIM, Oh Cheol
  • 정해영 JEONG, Haeyoung
Agents
  • 최규환 CHOI, Kyu Whan
Priority Data
10-2012-003823713.04.2012KR
10-2013-000222708.01.2013KR
Publication Language Korean (KO)
Filing Language Korean (KO)
Designated States
Title
(EN) METHOD FOR PRODUCING A TOXIC PROTEIN BY USING LACI PROTEIN MUTANT
(FR) PROCÉDÉ DE PRODUCTION D'UNE PROTÉINE TOXIQUE AU MOYEN D'UNE PROTÉINE LACI MUTANTE
(KO) LACI 단백질 변이체를 이용한 독성 단백질의 생산 방법
Abstract
(EN)
The present invention relates to a method for producing a toxic protein by using LacI protein mutant, a gene coding LacI protein mutant, a recombination vector including the gene, a host cell and the host cell transformed into the recombination vector, which controls precisely the expression of LacI mutant gene depending on the concentration of Rhamnose by using Rha promoter controlled by Rhamnose, so as to optimize the expression of the target protein to produce in a coliform bacillus cell, thus increasing productivity, and having great usefulness for the expression of a recombination toxic protein in a host cell including coliform bacillus.
(FR)
La présente invention concerne un procédé de production d'une protéine toxique au moyen d'une protéine LacI mutante, un gène codant une protéine LacI mutante, un vecteur de recombinaison contenant le gène, une cellule hôte et la cellule hôte transformée en vecteur de recombinaison. L'expression du gène LacI mutant est régulée avec précision en fonction de la concentration en rhamnose par le biais du promoteur Rha régulé par le rhamnose, ce qui permet d'optimiser l'expression de la protéine cible produite dans une cellule de colibacille et d'augmenter ainsi la productivité. L'invention est très utile pour l'expression d'une protéine toxique recombinée dans une cellule hôte, telle qu'un colibacille.
(KO)
본 발명은 LacI 단백질 변이체, 상기 LacI 단백질 변이체를 코딩하는 유전자, 상기 유전자를 포함하는 재조합 벡터, 상기 재조합 벡터로 형질전환된 숙주세포 및 상기 숙주세포를 이용하여 독성 단백질을 생산하는 방법에 관한 것으로, lacI 변이 유전자의 발현을 람노스에 의해 조절되는 Rha 프로모터를 이용하여 람노스의 농도 의존적으로 정밀하게 조절하며, 이에 의해 대장균 세포 내에서 생산하고자 하는 목적 단백질의 발현을 최적화함으로써 그 생산량을 증대시킬 수 있으므로, 대장균을 포함한 숙주세포에서 재조합 독성 단백질 발현에 그 유용성이 매우 크다.
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