以绿色荧光蛋白为报告基因的原核启动子检测体系构建  

Construction of Prokaryotic Promoter Function Analysis System Using Green Fluorescent Protein Report System

作　　者：陈坤[1] 黎明[1] 高伟[1] 路福平[1] 

机构地区：[1]天津市工业微生物重点实验室,天津科技大学生物工程学院,天津300457

出　　处：《生物技术通报》2008年第2期184-187,共4页Biotechnology Bulletin

基　　金：国家“863”计划资助项目(2007AA02Z212)

摘　　要：以质粒pMUTIN-GFP+扩增获得的gfp+基因为报告基因,将其克隆到大肠杆菌-枯草芽孢杆菌穿梭载体pBE2,构建成一个具有启动子活性检测功能的重组质粒pBE2-GFP+。将组成型启动子P43和诱导型启动子Pspac克隆入pBE2-GFP+,得到重组表达载体pBE-GFP-P43和pBE-GFP-Pspac,转化至大肠杆菌和枯草芽孢杆菌。荧光显微镜检测GFP+蛋白的表达情况。结果表明,2种不同类型的启动子均能在大肠杆菌BL21和枯草芽孢杆菌1A751中启动gfp+基因的表达。The gfp gene sequence was amplified from pMUTIN-GFP＋ by PCR. By recombination of the gfp PCR product and E.coli-B. subtilis shuttle vector pBE2,the vector pBE2-GFP＋ fuctioned as promoter analysis was constructed. In this study,we constructed expression vector pBE-GFP-P43 and pBE-GFP-Pspac by cloning the constitutive promoter P43 and inductive promoter Pspac into pBE2-GFP＋,respectively. GFP＋ protein was examined through fluorescent microscopy. It is indicated that the two kinds of promoters successfully drove the expression of gfp＋ gene in both E. coil BL21and B. subtilis1A751.

关 键 词：绿色荧光蛋白(GFP+) 启动子活性检测 大肠杆菌-枯草芽孢杆菌穿梭载体 

分 类 号：Q78[生物学—分子生物学] S668.108[农业科学—果树学]