生物加工过程2024，Vol.22Issue(2)：147-155,9.DOI:10.3969/j.issn.1672-3678.2024.02.004
停滞棒杆菌遗传改造工具和启动子文库构建
Genetic modification tools and synthetic promoter library for Corynebacterium stationis
摘要
Abstract
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.关键词
停滞棒杆菌/转化效率/穿梭质粒/启动子/基因敲除Key words
Corynebacterium stationis/transformation efficiency/shuttle plasmid/promoter/gene knockout分类
生物科学引用本文复制引用
叶光正,余心宇,林钰宽,蔡友华,邢新会,陈武,薛正莲,张翀..停滞棒杆菌遗传改造工具和启动子文库构建[J].生物加工过程,2024,22(2):147-155,9.基金项目
国家自然科学基金(21938004) （21938004）
