鸡贫血病毒VP1和VP2蛋白在家蚕中的联合表达

将鸡贫血病毒VP1和VP2基因分别克隆入转换载体pBacPAK8中，获得重组转移质粒pBac-vp1和pBac-vp2。以上两质粒分别与CvnⅠ酶切线性化的亲本病毒Bm-BacPAK6DNA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 BmNPV）是很有前途的CAV亚单位疫苗的生产系统。

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

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.