生物技术通报2024，Vol.40Issue(7)：108-116,9.DOI:10.13560/j.cnki.biotech.bull.1985.2024-0174
一种谷氨酸棒杆菌4-异丙基苯甲酸诱导型启动子的设计与应用
Design and Application of a Cumate-inducible Promoter for Corynebacterium glutamicum
摘要
Abstract
[Objective]Corynebacterium glutamicum is an important industrial microorganism,and its metabolic engineering modification by gene editing may effectively broaden the diversity of its fermentation products.The lack of high-intensity,low-leakage,and low-cost inducible promoters limits the metabolic engineering modification,thereby the design and application of novel inducible promoters are necessary.[Method]An cumate-induced promoter PH10-CuO was constructed by embedding the operator sequence CuO on the constitutive promoter PH10.[Result]Using green fluorescent protein as a quantitative reporter,the relative fluorescence intensity was low due to the basal leakage of PH10-CuO when without the inducer 4-isopropylbenzoic acid;and gfp expression significantly increased when the fluorescence intensity was up to 62 000 at presence of 25 μg/mL 4-isopropylbenzoic acid for 12 h.This proved that PH 10-CuO has very good rigor and induced expression intensity.Meanwhile,in Constructing the gene editing plasmid for C.glutamicum with PH10-CuO regulating the expression of recET and cas12a,The accurate editing of target gene and insertion of foreign genes in C.glutamicum chromosome.[Conclusion]The inducible promoter PH10-CuO presented the advantages with high intensity and low leakage,which enables it as an useful element to regulate temporal expression of specific genes in C.glutamicum.关键词
谷氨酸棒杆菌/诱导型启动子/阻遏蛋白CymR/4-异丙基苯甲酸Key words
Corynebacterium glutamicum/inducible promoter/repressor protein CymR/4-isopropylbenzoic acid引用本文复制引用
毋舒宁,苏永平,李冬雪,柏映国,刘波,张志伟..一种谷氨酸棒杆菌4-异丙基苯甲酸诱导型启动子的设计与应用[J].生物技术通报,2024,40(7):108-116,9.基金项目
国家重点研发计划(2022YFD1300703),国家自然科学基金项目(32171806) （2022YFD1300703）
