运动发酵单孢菌ZM4 adhⅡ基因的克隆及在大肠杆菌TOP10中的表达  

Cloning of Alcohol Dehydrogenase Ⅱ Gene from Zymomonas mobilis and Expression in Escherichia coli TOP10

作　　者：谭力[1] 陈介南[1] 张伟涛[1] 王义强[1] 何钢[1] 

机构地区：[1]中南林业科技大学生物环境科学与技术研究所,长沙410004

出　　处：《生物技术通报》2009年第1期87-90,94,共5页Biotechnology Bulletin

基　　金：国家948项目(2006-4-123)

摘　　要：利用高保真聚合酶从运动发酵单胞菌中克隆出乙醇脱氢酶基因,加A后克隆到pGM-T载体,测序验证无误。经酶切、酶连到表达载体pUC-18。形成重组质粒pUC-18-adhⅡ,转化到大肠杆菌TOP10中,经定性定量分析乙醇脱氢酶酶基因在大肠杆菌中高效表达,成功构建出乙醇脱氢酶基因的表达载体。The genes encoding alcohol dehydrogenase Ⅱ of Zymomonas mobilis was amplified by using high-fidelity DNA Polymerase from total DNA,and cloned into pGM-T vector after added A. The vector was verified by sequencing. The recombinant plasmid PUC18-adh Ⅱ was constructed by inserting expression vector pUC-18 after endonucleases digesting and ligating,and then transformed into E.coli TOP10. The recombinant strains were induced by IPTG to beexpressed. A/cohol dehydrogenasellgene was expressed highly by means of quantitive-qualitative analysis,and expression vector PUC18-adh Ⅱ was constructed in E.eoli TOP10.

关 键 词：运动发酵单胞菌 乙醇脱氢酶基因 克隆 

分 类 号：Q93[生物学—微生物学]