鸡贫血病毒VP1和VP2蛋白在家蚕中的联合表达  

Simultaneous Expression of Chicken Anaemia Virus Proteins VP1 and VP2 in Silkworms

作　　者：黄建芳[1] 张知良[1] 赵宝华[1] 胡清海[1] 肖庆利[2] 张志芳[2] 何家禄[2] 

机构地区：[1]江苏省家禽科学研究所,江苏扬州225003 [2]中国农业科学院蚕业研究所农业部家蚕生物技术重点开放实验室,江苏镇江211018

出　　处：《动物医学进展》2002年第3期53-56,共4页Progress In Veterinary Medicine

基　　金：江苏省"九五"农业科技攻关计划项目 (BE96 4 5 7)

摘　　要：将鸡贫血病毒 VP1和 VP2基因分别克隆入转移载体 p Bac PAK8中 ,获得重组转移质粒 p Bac-vp1和 p Bac-vp2。以上两质粒分别与Cvn 酶切线性化的亲本病毒 Bm-Bac PAK6DNA共转染家蚕细胞 ,通过蓝白斑筛选 ,纯化得到重组病毒 Bm-vp1和 Bm-vp2。PCR分析表明 Vp1和 Vp2基因已整合进杆状病毒基因组中。将 Bm-vp1和 Bm-vp2共感染 5龄家蚕 ,通过表达产物免疫 SPF鸡产生的抗血清与 CAV感染的 MDCC-MSB1细胞的间接荧光抗体分析 ,证明表达产物能诱导鸡产生相应的抗体。该研究表明 ,表达 VP1和 VP2蛋白的重组家蚕杆状病毒 ( recombinant Bm NPV)VP1 and VP2 genes of chicken anaemia virus were cloned into transfer vector pBacPAK8, then recombinant transfer plasmids pBac vp1 and pBac vp2 were obtained, respectively. The above two plasmids and parental baculovirus Bm BacPAK6 DNA (digested with Cvn I) were used to cotransfect BmN cells, respectively. Recombinant viruses Bm vp1 and Bm vp2 were selected and purified by blue white plaque assay. Vp1 and vp2 genes were integrated into baculovirus genomes by PCR analysis. Silkworms were co infected with Bm vp1 and Bm vp2. Antisera were obtained from SPF chickens inoculated with recombinant proteins. MDCC MSB1 cells infected with CAV reacted with the above antisera via immunofluorescence assay. The result showed the expression products could induce the corresponding antibody in chickens. Recombinant BmNPV expressing VP1 and VP2 is, therefore, a great hopeful production system for a subunit vaccine against CAV infection.

关 键 词：鸡贫血病毒 VP1 VP2蛋白 家蚕 联合表达 

分 类 号：S852.659.2[农业科学—基础兽医学]