PRRS病毒E基因的克隆及表达载体的构建  

Cloning of E Gene of PRRS Virus and Construction of Its Expressing Vectors

作　　者：苑博华[1] 蒋继志[1] 刘红梅[1] 

机构地区：[1]河北大学生命科学学院,河北保定071002

出　　处：《华北农学报》2006年第1期27-30,共4页Acta Agriculturae Boreali-Sinica

基　　金：河北省自然科学基金项目(300081);河北省省级重点学科生物工程

摘　　要：分别以质粒pBV221和pPIC9K为表达载体,成功构建了PRRS病毒E基因克隆载体,转化后经抗性筛选、PCR扩增、双酶切鉴定,确认得到了含有目的基因的阳性克隆。对含有E蛋白原核表达载体的阳性克隆进行诱导表达,经过酶联免疫鉴定,结果显示外源蛋白在宿主菌中有表达,成功实现了在大肠杆菌细胞中的克隆。本研究成功构建了PRRSV BJ-4毒株E基因的原核表达载体和真核表达载体,为基因工程疫苗的研制奠定了基础。The E gene fragments of PRRS strain BJ-4 have been amplified and cloned into plasmid pBV221 and pPIC9K. Recombinant plasmid was constructed and transformed into the JM109. The result indicated that the recombinant vectors were successfully constructed. Positive clones of target genes were screened by PCR and identified with restriction endo-nuclease analysis. The positive clone of recombinant prokaryotic expression vector was induced and ELISA showed that it could react with the antibody to PRRSV. The results showed that E Protein of PRRSV expressed with was expressed in prokaryote system.The prokaryotic expression vector and eucaryotic expression vector were successfully constructed, which would be useful for production of gene vaccination in future.

关 键 词：PRRSV 基因克隆 原核表达 真核表达 

分 类 号：Q342[生物学—遗传学]