军团菌MOMPS基因的克隆并在原核系统中表达  

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

作　　者：张雷[1] 陈建平[1] 王涛[1] 张莉[1] 田玉[1] 

机构地区：[1]四川大学华西基础医学与法医学院寄生虫学教研室形态学实验室,成都610041

出　　处：《生物医学工程学杂志》2006年第2期379-382,共4页Journal of Biomedical Engineering

基　　金：国家自然科学基金资助课题(39870656)

摘　　要：以军团菌DNA为模板,PCR扩增获得军团菌主要外膜蛋白基因(M a jor ou ter m em brane prote in gene,om pS),与原核表达质粒pUC 18定向重组,构建重组质粒,转化大肠杆菌BL 21,并用限制性酶酶切分析、聚合酶链式反应、核酸序列分析、十二烷基磺酸钠-聚丙烯酰胺凝胶电泳、W estern印迹进行鉴定。实验结果表明我们扩增出了军团菌914 bp的om pS基因,成功构建了重组质粒pLPom pS,并在原核系统中得到了表达。In this study, the ompS gene, a major outer membrane protein gene of Legionella pneumophila, was obtained from the DNA of Legionella pneumophila by PCR. The gene was cloned into prokaryotic expressional plasmid pUC18 to construct recombinant plasmid. The recombinant plasmid was transformed into E. coli strain BL21. The identification was made by means of restriction enzyme analysis, polymerase chain reaction, DNA sequencing analysis, SDS - polyacrylamine gel electrophoresis analysis and Western blot. The results showed that the ompS gene of 914 bp was amplified from Legionella pneumophila DNA, the recombinant plasmid pLPompS was constructed and its expression in prokaryotic cell was detected successfully.

关 键 词：军团菌 主要外膜蛋白基因(ompS) 克隆 表达 

分 类 号：R346[医药卫生—基础医学]