耻垢分枝杆菌MSMEG_6935基因的敲除对其生物被膜产量的影响

Effect of MSMEG_6935 deletion mutant on biofilm production in Mycobacterium smegmatis

探索出一套分枝杆菌基因敲除的可靠方案,并研究耻垢分枝杆菌MSMEG_6935基因对其生物被膜产量的影响,结核分枝杆菌相关基因的敲除对其致病机理的深入研究有重要意义。利用融合PCR方法将MSMEG_6935基因的上、下游基因以及替代目的基因的kan基因融合,并将产物克隆到温敏型穿梭质粒pPR27,再电转入耻垢分枝杆菌mc2155中,筛选获得MSMEG_6935基因缺失株mc2155-6935。进一步研究MSMEG_6935基因的敲除对细菌生长曲线和生物被膜产量的研究。成功构建了MSMEG_6935基因的缺失株,并研究证明耻垢分枝杆菌MSMEG_6935基因的缺失对耻垢分枝杆菌的生长曲线没有影响,但会对其生物被膜的产量产生较大影响。

To explore a reliable gene knockout method in Mycobacteria,and research the effect of MSMEG_6935deletion mutant on biofilm production in Mycobacteriumsmegmatis.The related gene knockout is important for Mycobacteria tuberculosis study.The upstream,downstream and kan flanking DNA fragments of target gene were fused by PCR,and then cloned into the shuttle vectorpPR27,the recombinant plasmid was introduced into mc2155 strain.In addition,we research the effect of MSMEG_6935deletion mutant on growth and biofilm production in Mycobacterium smegatis.The MSMEG_6935gene deletion strains were successfully constructed and the MSMEG_6935gene knockout has no effect on the growth curve of mc2155,but it has a great effect on biofilm production.