单核细胞增生性李斯特菌iap基因的原核表达  

Prokaryotic Expression of iap Gene from Listeria monocytogenes

作　　者：区燕宜[1] 徐成刚[1] 叶贺佳[1] 吴晓薇[1] 廖明[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642

出　　处：《华南农业大学学报》2008年第2期108-111,共4页Journal of South China Agricultural University

基　　金：广东省自然科学基金创新团队项目(5200638)

摘　　要：该研究利用自行设计的引物通过PCR方法扩增出单核细胞增生性李斯特菌Listeria monocytogenes C5305株的iap基因，在iap基因的5’端和3’端分别引入EcoRⅠ和XhoⅠ2个酶切位点并将其克隆到pET-28a表达载体上，经PCR鉴定、酶切鉴定和核苷酸序列测定后获得重组质粒pET-28a—iap，将该重组质粒转化入大肠杆菌B121（DE3）pLysS，经1mmol／LIPTG诱导4—6h后，通过SDS—PAGE及Westem-blotting鉴定，证明该基因得到表达，表达产物相对分子质量约为60000，以可溶形式存在．In this study, the iap gene of Listeria monocytogene strain C5305 was amplified by PCR using specific primers. EcoR Ⅰ and XhoⅠ sites were induced to 5＇ and 3＇ end of iap gene, respectively. Specific PCR products were then cloned into expression vector plasmid pET-28a, and recombinant plasmid ： pET-28a-iap was verified by PCR, restriction enzyme digestion and sequencing. After transforming pET- 28a-iap into E. coli BL21 （DE3）pLysS and inducing with 1 mmol/L IPTG for4 -6 h, the iap gene expressed successfully in soluble form and with a relative molecular mass of approximately 60 000.

关 键 词：单核细胞增生性李斯特菌 iap基因 原核表达 

分 类 号：S852.6[农业科学—基础兽医学]