停滞棒杆菌遗传改造工具和启动子文库构建OACSTPCD

Corynebacterium stationis(C.sta)has been widely utilized in the industrial fermentation production of inosine 5'-monophosphate(IMP).Both genetic modification tools and synthetic promoter libraries are quite important in the optimization of cell factories using metabolic engineering approaches.Herein,we developed a stable expression system for C.sta heterologous genes.Upon the formation of a novel E.coli-C.sta shuttle vector based on the pCG1 replicon,an exogenous DNA transformation technique was developed for C.sta.Meanwhile,a gene knockout method was established through homologous recombination of suicide plasmid.As a result,the electro-transformation efficiency of C.sta could reach(2.3±0.3)×103cfu/μg DNA.Furthermore,a synthetic promoter library was constructed with 24 promoters across the range of 30-fold strength difference,which could be potentially used in the fine regulation of gene expression in C.sta.