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运动发酵单孢菌ZM4-pdc基因的克隆及在大肠杆菌TOP10中的表达与活性分析 被引量:1

Cloing and Expression Activity Analysis of pdc Gene from Zymomonas mobilis ZM4 in E.coli TOP10
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摘要 利用高保真聚合酶从运动发酵单胞菌中克隆出丙酮酸脱羧酶基因,加A后克隆到pGM-T载体,测序验证无误。经酶切、连接、转化到表达载体pUC-18。形成重组质粒pUC-18-pdc,转化到大肠杆菌TOP10中,经定性定量分析丙酮酸脱羧酶基因在大肠杆菌中高效表达,成功构建出乙醛的代谢途径。 The gene pdc encoding pyruvate decarboxylase of Zymomonas mobilis was amplified using High- fidelity DNA Polymerase ftom total DNA. Cloned into pGM-T vector after adding A. The vector was verified by sequencing. The recombinant plasmid,pUC-18-pdc was constructed by inserting pdc into expression vector pUC-18 after end onucleases digesting and ligating,and then transformed E. coliTOP10. The recombinant strains were induced by IPTG to express pyruvate decarboxylase is expressing highly by means of quantitive-qualitative analysis,Aldehyde metabolism pathway is constructed in E.coliTOP10.
出处 《生物技术通报》 CAS CSCD 2008年第2期151-154,190,共5页 Biotechnology Bulletin
基金 国家948项目(2006-4-123)
关键词 运动发酵单孢菌 丙酮酸脱羧酶 克隆 乙醛 代谢工程 Zymornonas rnobilis Pyruvate decarboxylase Cloning Aldehyde Metabolic engineering
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