表达Ⅰ型鸭病毒性肝炎病毒VP1蛋白重组腺病毒的构建  

Construction of recombinant adenovirus expressing VP1 protein of duck viral hepatitis virus Ⅰ

作　　者：殷秀辰[1] 刘红玉[2] 陈玉环[2] 刘明[1] 张云[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所,黑龙江哈尔滨150001 [2]东北农业大学动物医学院,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2013年第7期526-529,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：现代农业水禽产业技术体系岗位科学家专项(CARS-43-10)

摘　　要：为构建表达Ⅰ型鸭病毒性肝炎病毒(DHV-Ⅰ)VP1蛋白的重组腺病毒,本实验采用RT-PCR方法扩增DHV-Ⅰ HP-1株VP1基因,克隆于腺病毒穿梭质粒pShuttle-CMV中,并将其电转化于含腺病毒pAdeasy骨架质粒的大肠杆菌BJ5183中,通过同源重组制备含有重组腺病毒的质粒。重组质粒经PacⅠ酶切线性化后转染AD293细胞,获得重组腺病毒(rAd-VP1)。间接免疫荧光和western blot鉴定结果显示,DHV-Ⅰ的VP1蛋白在重组腺病毒感染的AD293中获得表达。rAd-VP1的制备为鸭免疫实验效果评价奠定了基础。In order to construct recombinant adenovirus expressing the VP1 protein of duck hepatitis virus type I （DHV-I）, the VPI gene was amplified by RT-PCR and cloned into pShuttle-CMV vector. The recombinant plasmid was transferred into E. co/i BJ5183 competent cells containing adenovirus backbone vector to produce recombinant adenovirus plasmid by homologous recombination. The positive recombinant plasmid was linearized by Pac I and transfected into AD293 cells. The recombinant adenovirus （rAd-VPI） was rescued and identified by indirect immunofluorescence assay and westem blot. The results showed that rAd-VPl expressing VPI protein of DHV- I was constructed successfully which laid the foundation for the evaluation of immune efficacy in ducks.

关 键 词：重组腺病毒 I型鸭病毒性肝炎 VP1蛋白 

分 类 号：S852.65[农业科学—基础兽医学]