军团菌外膜蛋白抗原基因的克隆并在原核系统中表达

Cloning of Major Outer Membrane Protein Antigen Gene of Legionella pneumophila and Detection of Its Expression in Prokaryotic Cell

目的 克隆军团菌外膜蛋白抗原基因 omp M,构建重组质粒 p LPM,并在原核系统中表达。方法 采用聚合酶链反应 (PCR)从军团菌基因组 DNA中扩增得到 omp M基因 ,导入载体 p UC1 8,在 JM1 0 9中表达 ,并用SDS-PAGE进行鉴定。结果 扩增出 773 bp的 omp M基因 ;构建重组质粒 p LPM;表达出 2 5 k Da的蛋白质。结论 成功扩增军团菌 omp M基因 ,构建重组质粒 p LPM,并在原核系统中得到了表达。

Objective To clone the major outer membrane antigen gene omp M of Legionella pneumophila, to construct recombinant plasmid pLPM and to dete ct its expre ssion in prokaryotic cell strain JM109.Methods The ompM gene fr o m Legionella pneumophila was amplified with PCR. The PCR product was cloned into pUC18 vector a nd ompM expressed in JM109. After that, the expression of JM109 of ompM gene was det ected with SDS-PAGE. Results The ompM gene of 773bp was amplif ied, the recombin ant plasmid pLPM constructed and protein of 25 kDa expressed in JM109.C onclusion The ompM gene was amplified, the recombinant plasmid pLPM cons tructed and its expression detected in prokaryotic cell successfully.