METHOD FOR QUANTITATIVELY CONTROLLING PLASMID COPY NUMBER IN ANTIBIOTIC-FREE PLASMID MAINTENANCE SYSTEM

WIPO Patent Application WO/2018/186704

A1

The present invention relates to a gene expression cassette comprising a synthetic 5' untranslated region (5' UTR), a promoter, and a regulatory gene; a recombinant vector comprising a replication origin and the gene expression cassette; a recombinant microorganism which has the recombinant vector introduced thereinto and shows alleviated segregational instability and ; a method for preparing a recombinant microorganism having alleviated segregational instability by introducing the recombinant vector thereinto; and a method for quantitatively controlling a plasmid copy number in a recombinant microorganism. According to the present invention, removal of infA and efp, which are genes indispensable for cells, coding respectively for a translation initiation factor and a protein elongation factor (EF-P), from a microbial chromosome and introduction of the gene expression cassette comprising the regulatory gene with Escherichia coli serving as a host allow the stable maintenance of plasmids in an antibiotic-free medium without causing intercellular intrinsic variations. In addition, the precise control of expression levels of infA and efp in the recombinant microorganism by means of a promoter can lead to the quantitative control of PCN at high yield as well. Therefore, the present invention can find a broad spectrum of applications in a variety of industries producing recombinant proteins.

WO/2006/130806	ASSAY FOR AGONISTS AND ANTAGONISTS OF ION CHANNELS AND FOR REGULATORS OF GENETIC EXPRESSION
WO/2007/035323	HYBRID PORTABLE ORIGIN OF REPLICATION PLASMIDS
JP2604478	[Title of Invention] Method for Producing Gene Product

JUNG GYOO YEOL (KR)
SEO SANG WOO (KR)
KANG CHAE WON (KR)
LIM HYUN GYU (KR)

PCT/KR2018/004038

October 11, 2018

April 05, 2018

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POSTECH ACAD IND FOUND (KR)
SEOUL NAT UNIV R&DB FOUNDATION (KR)

C12N15/69; C12N15/70; C12P5/02; C12P7/46

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KIM, Soon Woong (KR)

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