谷氨酸棒杆菌argG基因缺失菌株的构建  

Construction of Corynebacterium glutamicum mutant with knockout of argG gene

作　　者：汤俊[1] 郝宁[1] 许晟[1] 许琳[1] 严明[1] 

机构地区：[1]南京工业大学生物与制药工程学院,江苏南京210009

出　　处：《南京工业大学学报（自然科学版）》2013年第6期86-90,共5页Journal of Nanjing Tech University(Natural Science Edition)

基　　金：江苏省普通高校研究生科研创新计划(CXLX11_0373)

摘　　要：构建1株谷氨酸棒杆菌argG基因缺失菌株并观察其对L-瓜氨酸生产的影响。采用crossover PCR方法扩增带有argG基因上下游同源序列的DNA片段,连接至载体pK18mobsacB,构建敲除载体pK18-ΔargG,转化至C.glutamicum ATCC 13032,利用卡那霉素抗性正筛选和果聚糖蔗糖sacB基因负筛选获得argG基因缺失菌株。发酵结果表明,argG缺失菌株60 h积累2.52 g/L L-瓜氨酸,出发菌C.glutamicum ATCC 13032不能积累L-瓜氨酸。Construction of argG-deleted mutant and effect of argG inactivation on L-citrulline production in Corynebacterium glutamicum were investigated. For deletion of argG gene in C. glutamicum ATCC 13032, crossover PCR was carried out to generate the argG deletion fragment ligated to the suicide vector pK18mobsacB. The recombinant plasmid pK18-AargG was transferred into C. glutamicum by eleetropo- ration. C. glutamicum ATCC 3032-△argG was successfully obtained by double homologous recombina- tion. The fermentation result showed that L-citrulline production of the argG gene knockout strain was achieved up to 2.52 g/L.

关 键 词：谷氨酸棒杆菌 L-瓜氨酸 基因敲除 

分 类 号：TQ922.9[轻工技术与工程—发酵工程]