A method for genome editing of yarrowia lipolytica based on homologous recombination

Abstract

This invention provides a genome editing method for *Yersinia lipolytica* based on homologous recombination, belonging to the fields of microbial genetic engineering and synthetic biology. This invention constructs the tool plasmid pUrloxP3 to achieve efficient knockout and combined knockout of ylAS and ylPDH, completing site-specific integration and functional verification of exogenous genes RgTAL, 4CL, and CHS. The Cre / loxP system is used to recover the URA3 marker and eliminate the pYLXP'-Cre helper plasmid. This invention features high editing efficiency, standardized procedures, cyclical selection markers, and stable integration of exogenous genes, ultimately yielding a stable engineered strain without markers or helper plasmids. It is suitable for multi-round metabolic engineering and cell factory construction of *Yersinia lipolytica*.

Images