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