以β-半乳糖苷酶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 β-galactosidase Report System

作　　者：陈坤[1] 黎明[1] 樊欣迎[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》2008年第1期13-16,共4页Biotechnology

基　　金：天津市科技发展计划项目资助(06YFGPSH03500)

摘　　要：目的:以β-半乳糖苷酶为报告基因构建原核启动子检测体系。方法:以由质粒pDL扩增获得的bgaB基因为报告基因,将其克隆到大肠杆菌-枯草杆菌穿梭载体pBE2,构建成一个具有启动子活性检测功能的重组质粒pBEB。将组成型启动子P43和诱导型启动子Pspac克隆入pBEB,得到重组表达载体pBEBP43和pBEBPspac,转化至大肠杆菌和枯草芽孢杆菌。结果:两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bgaB基因的表达。结论:成功构建具有较为广泛适用性的原核启动子检测体系。Objective： Construction of prokatyotic promoter function analysis system using β- galactosidase report system. Methods： The bgaB gene sequence was amplified from pDL by PCR. The vector pBEB fuctioned as promoter analysis was constructed by recombination of the bgaB PCR product and E. coli - B. subtilis shuttle vector pBE2.In this study, we constructed expression vector pBEBP43 and pBEBPapac by cloning the constitutive promoter P43 and inductive promoter Papac into pBEB, respectively. Results： The two kinds of promoters successfully drove the expression of BgaB protein in both E. coli BL21 and B. subtilis 1A751. Conclusion： A useful prokaryotic promoter function analysis vector was obtained.

关 键 词：Β-半乳糖苷酶 启动子活性检测 大肠杆菌-枯草芽孢杆菌穿梭载体 

分 类 号：Q786[生物学—分子生物学]