猪传染性胃肠炎病毒S基因A、D抗原共表达的复制缺陷型重组腺病毒构建及鉴定  

Construction and Identification of Replication defective Recombinant Adenovirus Expressing A and D epitope of S protein gene in Transmissible gastroenteritis virus

作　　者：刘文晓 高志强[2] 蒲静[2] 张伟[2] 刘环[2] 牛建蕊 张鹤晓[2] 

机构地区：[1]北京森康生物技术开发有限公司,北京怀柔101400 [2]北京出入境检验检疫局,北京朝阳100026

出　　处：《中国兽医杂志》2016年第9期3-6,共4页Chinese Journal of Veterinary Medicine

基　　金：北京市优秀人才培养资助青年骨干个人项目(2015-000097607G290)

摘　　要：通过RT-PCR技术和重叠PCR技术扩增出含有猪传染性胃肠炎病毒S基因的A抗原表位和D抗原表位,构建复制缺陷型腺病毒穿梭质粒。穿梭质粒与腺病毒骨架质粒经PacI线性化之后,共同转染AD-293细胞,获得共表达A、D抗原表位的复制缺陷型重组腺病毒。经Western Blotting检测,该重组腺病毒能够正确表达目的蛋白基因,而且目的蛋白能够与猪传染性胃肠炎阳性血清反应。本研究结果为猪传染性胃肠炎重组疫苗研发奠定基础。RT-PCR and overlap PCR were used to amplify the A and D epitope of spikes protein（S protein） gene in Transmissible gastroenteritis virus（TGEV）.The A and D epitope genes were introduced into the shuttle vector.To generate the recombinant adenovirus,the shuttle vector containing the gene of interest and the recombinant adenoviral backbone were co-transfected into AD-293 cells.RT-PCR was applied to detect the genes of A and D proteins in the recombinant adenovirus.The Western blotting results revealed that the A and D genes were properly expressed,and the expressed protein could react with TGEV-positive serum.The study sets a foundation for the preparation of recombinant vaccine to control TGEV infection.

关 键 词：猪传染性胃肠炎 复制缺陷型腺病毒 疫苗 

分 类 号：S855.3[农业科学—临床兽医学]