变形链球菌recA基因启动子荧光蛋白表达载体的构建  

Construction of a red fluorescent shuttle vector controlled by recA operon promoter of Streptococcus mutans

作　　者：黄文明[1] 徐仰龙[1] 杨德琴[1] 

机构地区：[1]遵义医学院附属口腔医院口腔内科,贵州563003

出　　处：《中华口腔医学杂志》2012年第5期291-295,共5页Chinese Journal of Stomatology

基　　金：基金项目：国家自然科学基金（30960419）;贵州省优秀科技教育人才省长专项基金[黔省专合字2005（244）号]

摘　　要：目的构建由recA基因启动子启动的红色荧光蛋白穿梭表达载体，研究变形链球菌recA基因表达的特点，以期为不同致龋环境中变形链球菌致龋毒力因子基因的表达提供参照。方法以变形链球菌（UA159）基因组DNA为模板，扩增recA基因启动子，采用双酶切反应连接入载体pDsRed2-N1，构建原核表达载体pRred；通过酶切重组入穿梭载体pDL276，构建红色荧光蛋白表达载体pLRred。结果经酶切鉴定目的质粒片段插入无误，同时重组载体pLRred转化菌荧光观测结果提示，重组载体pLRred构建成功。结论本项研究中构建的recA基因启动子红色荧光蛋白同源重组克隆载体正确，可应用于recA基因表达的研究，同时可为研究变形链球菌致龋基因的表达提供内参照。Objective To construct a red fluorescent shuttle vector controlled by recA operon promoter to transform Streptococcus mutans. Methods The promoter of recA was amplified from Streptococcus mutans UA159, and connected to plasmid pDsRed2-N1 to construct pRred with a red fluorescent coding gene, which was then inserted into the shuttle vector pDL276 to construct pLRred. Results pLRred was successfully constructed, and Escherichia coli transformed with the pLRred plasmid could express reporter gene DsRed. Conclusions The recombination plasmid pLRred can be used in the further research of the expression of cariogenic virulence factor gene by Streptococcus mutans in biofilm.

关 键 词：链球菌 变异 Rec A重组酶 红色荧光蛋白 管家基因 

分 类 号：Q78[生物学—分子生物学]