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中国农学通报

中国农学通报 ›› 2005, Vol. 21 ›› Issue (5): 20-20.

所属专题： 生物技术； 畜牧兽医

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逆转录病毒载体介导的猪瘟病毒E2基因在真核细胞中表达及动物免疫试验

许信刚,胡建和,张彦明   

  • 出版日期:2005-05-05 发布日期:2005-05-05

Retroviral Vector Mediated CSFV E2 Envelope Protein Gene Expression Eukaryotic Cells and Animal Immune Experiment

Xu Xingang, Hu Jianhe, Zhang Yanming   

  • Online:2005-05-05 Published:2005-05-05

摘要： 利用DNA重组技术将猪瘟病毒(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，该重组逆转录病毒表达载体与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SP2/0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CSFV E2基因在SP2/0细胞膜上成功表达。将表达E2蛋白的SP2/0细胞腹腔免疫BALB/c小鼠，用流式细胞仪检测证明，成功诱导小鼠产生了抗E2蛋白的抗体。为下一步利用细胞免疫小鼠研制抗猪瘟病毒单克隆抗体打下基础。

Abstract: 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 cells 293GP by calcium phosphate transfection method thus the pseudovirus were producesd . The pseudovirus infected eukaryotic cells SP2/0 and expression E2 protein was determined by puromycin-resistant and FACS analysis. BALB/c mice were intraperitoneal injected with SP2/0 cells which expressed CSFV E2 protein. The result showed anti-CSFV E2 antibody was screened by FACS.