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中国农学通报 ›› 2005, Vol. 21 ›› Issue (3): 9-9.

所属专题: 生物技术 畜牧兽医

• 目次 • 上一篇    下一篇

猪瘟病毒DNA疫苗的构建及动物免疫试验

许信刚,胡建和 ,张彦明   

  • 出版日期:2005-03-05 发布日期:2005-03-05

Construction of CSFV E2 Envelope Protein Gene Vaccine and Animal Immune Experiment

Xu Xingang, Hu Jianhe, Zhang Yanming   

  • Online:2005-03-05 Published:2005-03-05

摘要: 利用DNA重组技术将猪瘟病毒(CSFV) C株E2囊膜蛋白全长基因克隆到真核表达载体pcDNA4.0的CMV启动子下游,采用磷酸钙转染法将重组质粒转入293T细胞,流式细胞仪(FACS)检测293T细胞瞬时表达了E2囊膜蛋白。将构建的重组质粒肌肉注射BALB/c小鼠,用流式细胞仪和酶联免疫吸附试验(ELISA)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DNA疫苗免疫小鼠研制抗猪瘟病毒单克隆抗体打下基础。

Abstract: CSFV gene vaccine was constructed by inserting full-length cDNA of CSFV Shimen strain E2 gene into an eukaryotic expression vector pcDNA4.0. The recombinant plasmid was transfected into eukaryotic cells 293T by calcium phosphate transfection method and transient expressive E2 envelope protein was analyzed by FACS. BALB/c mice were intramuscular injection with the recombinant plasmid. Anti-CSFV E2 antibody was screened by FACS and ELISA. The results showed that CSFV E2 protein was expressed transciently in 293T cells. Specific anti-E2 antibody could be detected in DNA immune mouse serum by FACS and ELISA..