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Position: Home > Articles > Cloning of Alcohol Dehydrogenase Ⅱ Gene from Zymomonas mobilis and Expression in Escherichia coli TOP10 Biotechnology Bulletin 2009,2009 (1) 87-90+94

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

作  者:
谭力;陈介南;张伟涛;王义强;何钢
单  位:
中南林业科技大学生物环境科学与技术研究所
关键词:
运动发酵单胞菌;乙醇脱氢酶基因;克隆
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。
译  名:
Cloning of Alcohol Dehydrogenase Ⅱ Gene from Zymomonas mobilis and Expression in Escherichia coli TOP10
作  者:
Tan Li Chen Jienan Zhang Weitao Wang Yiqiang He Gang (Institute of Biological and Enviromental Science & Technology,Central South University of Forestry and Technology, Changsha 410004)
关键词:
Zymomonas mobilis Pyruvate decarboxylase Cloning
摘  要:
The genes encoding alcohol dehydrogenaseⅡof 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. Alcohol dehydrogenaseⅡgene was expressed highly by means of quantitive-qualitative analysis,and expression vector PUC18-adhⅡ was constructed in E.coli TOP10.

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