逆转录病毒载体介导的猪瘟病毒E2基因的真核表达  

Expression of CSFV E2 Envelope Protein Gene in Eukaryotic Cells and Preparation of Monoclonal Antibodies against CSFV E2 Envelope Protein

作　　者：刘建玲[1] 张彦明[1] 苏正元[1] 许信刚[1] 

机构地区：[1]西北农林科技大学动物科技学院

出　　处：《中国病毒学》2006年第3期249-252,共4页Virologica Sinica

基　　金：陕西省科技攻关项目(2003K02-G11-01);陕西省教育厅专项科研计划项目(05JK296)

摘　　要：利用DNA重组技术将猪瘟病毒(Classicalswinefevervirus,CSFV)石门株囊膜蛋白E2基因插入逆转录病毒载体pBABE-puro中构建成重组逆转录病毒载体pBABE-puro-E2,该重组逆转录病毒载体与pVSVg质粒经磷酸钙共转染法将其转入293GP细胞中包装逆转录病毒假病毒。用包装的假病毒感染PK-15细胞,经嘌呤霉素筛选阳性细胞后进行流式细胞技术(FACS)分析,结果表明CSFVE2基因在PK-15细胞膜上成功表达。将表达E2蛋白的PK-15细胞腹腔免疫Balb/c小鼠,成功诱导小鼠产生了抗E2蛋白的抗体。The recombinant retroviral vector pBABE-puro-E2 was constructed by inserting full-length cDNA of CSFV Shimen strain E2 gene into pBABE-puro. Both the recombinant retroviral vector and pVSVg plasmid were transfected into eukaryotic ceils 293GP by calcium phosphate transfection method, and the pseudovirus were produced. The pseudovirus-infected eukaryotic cells PK-15 and expression of E2 protein were determined by puromycin-resistant and FACS analysis. Balb/c mice were intraperitoneal injected with PK-15 cells expressing the CSFV E2 protein. Anti-CSFV E2 antibody was screened by ELISA. The results showed that CSFV E2 protein was expressed in PK-15 cells＇ envelope protein successfully. ELISA could detect specific anti-E2 antibody in mouse serum immuned by PK-15 cells.

关 键 词：猪瘟病毒 E2囊膜蛋白 逆转录病毒载体 FACS 

分 类 号：S852.65[农业科学—基础兽医学]