鸡传染性贫血病毒vp3基因在真核细胞中的表达与功能分析  

The cloning and eukaryotic expression of vp3 gene and its functional analysis

作　　者：黄超华[1,2] 张全红[3] 陈坤[1] 王永强[1] 曹红[1] 李晓齐[1] 郑世军[1] 

机构地区：[1]中国农业大学动物医学院,北京海淀100193 [2]蛇口出入境检验检疫局,广东深圳518054 [3]国家知识产权局专利局专利审查协作北京中心,北京海淀100190

出　　处：《中国兽医杂志》2015年第7期7-10,共4页Chinese Journal of Veterinary Medicine

基　　金：国家自然科学基金项目(31272543);现代农业产业技术体系建设专项资金(NYCYTX-41)

摘　　要：为研究鸡传染性贫血病毒(CIAV)VP3蛋白在宿主细胞中的作用,本研究采用PCR技术从CIAV DNA中扩增出vp3基因,并将该基因克隆到p MD18-T simple vector中。经测序证实所克隆的vp3基因与CIAV CUX-1标准株相同。并将vp3基因亚克隆到真核表达载体p EGFP-N1中,构建了重组真核表达质粒p EGFP-vp3与p CMV-Myc vp3,转染BHK-21和293 T细胞,经RT-PCR、荧光显微镜以及Western Blot证实转染的真核细胞能够表达VP3蛋白。以台盼蓝染色法检测细胞死亡,结果表明,VP3蛋白具有较强诱导细胞死亡的作用。该结果为深入研究CIAV的致病机理奠定了基础。In order to elucidate the role of chicken infectious anemia virus（CIAV）VP3 protein in host cells,vp3 gene was cloned from the viral DNA by PCR and was inserted into the p MD18-T simple vector. The result of DNA sequencing showed that the sequence of vp3 gene was the same as that of CUX-1.The vp3 gene was then sub-cloned into the eukaryotic expression vector p EGFP-N1 or p CMV-Myc.The p EGFP-vp3 or p CMV-Myc-vp3 plasmids were transfected into the BHK-21 or 293 T cells for the expression of VP3.The expression of VP3 by the transfected cells was determined by RT-PCR,fluorescence microscope and Western Blot.VP3-induced cell death was measured by the Trypan Blue-staining.The results showed that the protein VP3 could be expressed in the transfected cells and induce cell death.These data lay a foundation for elucidation of pathogenesis of CIAV.

关 键 词：鸡传染性贫血病毒 VP3基因 凋亡素 真核表达 

分 类 号：S852.65[农业科学—基础兽医学]