羊布鲁菌16M Omp25真核表达载体的构建  

The construction of Brucella melitensis 16M Omp25 gene in eukaryotic expression vector

作　　者：屈海龙[1,2] 王海浪[3] 陈思[1,2] 张瑞[1,2] 王秀然[1,2] 钱晶[1,2] 王兴龙[1,2] 

机构地区：[1]军事医学科学院军事兽医研究所,长春130122 [2]吉林省人兽共患病预防与控制重点实验室,长春130122 [3]北京奶牛中心,北京100192

出　　处：《中国兽医学报》2013年第12期1848-1850,1854,共4页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30972198/C180503);国家科技支撑计划(2010BAD04B03)

摘　　要：利用聚合酶链式反应(PCR),以羊布鲁菌16M基因组为模板克隆Omp25基因,设计含有EcoRⅠ和XhoⅠ酶切位点的引物序列,定向克隆到真核表达载体pCMV-HA上,构建真核表达重组质粒pCMV-HA-Omp25。经测序及双酶切验证正确,证明成功构建了pCMV-HA-Omp25真核表达质粒,为表达纯化外膜蛋白Omp25,研究其功能奠定基础。In this research,Brucella rnelitensis 16M Omp25 gene was amplified by PCR. The puri- fied PCR products were ligated with pMD18-T simple vector. After enzyme digestion and sequence analysis,the correct pMD-18T-Omp25 gene was digested by EcoR I and Xho,and cloned into the eukaryotic expression vector pCMV-HA which was digested by the same enzymes. The result shows that the eukaryotic expression vector of Brucella rnelitensis 16M Omp25 gene was con- structed successfully,which laid the foundation for the Omp25 function analysis.

关 键 词：羊布鲁菌16M Omp25 真核表达 

分 类 号：S852.61[农业科学—基础兽医学]