CAJ | 학술논문

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;通过酶切重组人穿梭载体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.
목적 구건유recA기인계동자계동적홍색형광단백천사표체재체,연구변형련구균recA기인표체적특점,이기위불동치우배경중변형련구균치우독력인자기인적표체제공삼조.방법 이변형련구균(UA159)기인조DNA위모판,확증recA기인계동자,채용쌍매절반응련접입재체pDsRed2-N1,구건원핵표체재체pRred;통과매절중조인천사재체pDL276,구건홍색형광단백표체재체pLRred.결과 경매절감정목적질립편단삽입무오,동시중조재체pLRred전화균형광관측결과제시,중조재체pLRred구건성공.결론 본항연구중구건적recA기인계동자홍색형광단백동원중조극륭재체정학,가응용우recA기인표체적연구,동시가위연구변형련구균치우기인적표체제공내삼조.
Objective To construct a red fluorescent shuttle vector controlled by recA operon promoter to transform Streptococcus mutaus.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.