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Construction of a grpE-based plasmid addiction system in Escherichia coli and its application in phloroglucinol biosynthesis

J Appl Microbiol. 2024-05; 
Ji-Ming Wang, Yu-Jin Cao, Xiao Men, Hai-Bo Zhang
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Proteins, Expression, Isolation and Analysis … A cloning plasmid pUC57S-grpEm carrying the synonymously re-encoded grpE expression cassette grpEm was purchased from Nanjing GenScript Biotech Co., Ltd. (Nanjing, China) (… Get A Quote

摘要

objective: Biotechnical processes in Escherichia coli often operate with artificial plasmids. However, these bioprocesses frequently encounter plasmid loss. To ensure stable expression of heterologous genes in E. coli BL21(DE3), a novel plasmid addiction system (PAS) was developed. results: This PAS employed an essential gene grpE encoding a cochaperone in the DnaK-DnaJ-GrpE chaperone system as the selection marker, which represented a chromosomal ΔgrpE mutant harboring episomal expression plasmids that carry supplementary grpE alleles to restore the deficiency. To demonstrate the feasibility of this system, it was implemented in phloroglucinol (PG) biosynthesis, manifesting improved host tolerance to PG and i... More

關鍵詞

grpE, essential gene, phloroglucinol, plasmid addiction system, tolerance, transcriptomic analysis
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