犬瘟热病毒V蛋白的表达、鉴定及亚细胞定位

Expression,Identification and Subcellular Localization of V Protein of Canine Distemper Virus

为研究犬瘟热病毒(canine distemper virus,CDV)V蛋白的功能,将CDVV基因片段与pGEX-6P-1载体连接,构建pGEX-6P-1-CDV-V重组表达质粒,通过原核表达系统表达了V蛋白,并将纯化的V蛋白免疫BALB/c小鼠,制备阳性血清。同时,将CDVV基因片段与pcDNA3.1载体连接,构建pcDNA3.1-CDV-V重组表达质粒,经转染Vero细胞后,用激光共聚焦技术确定V蛋白在真核细胞中的分布及亚细胞定位。结果显示,成功表达了V蛋白,并制备了阳性血清。重组质粒pcDNA3.1-CDV-V在外源真核细胞Vero中获得了表达,表达蛋白主要聚集于细胞质。本试验结果为进行CDV V蛋白的功能研究奠定了基础。

In order to study the function of canine distemper virus （CDV）V protein,V gene frag-ment was inserted into the pGEX-6p-1 vector,and then pGEX-6p-1-CDV-V recombinant expres-sion plasmid was obtained.Purified V protein immunized BALB/c mice to prepare the positive se-rum.Meanwhile,to confirm the distribution of V protein and subcellular localization with confocal laser technology,a eukaryotic expression recombinant plasmid pcDNA3.1-CDV-V was built,then transfected into Vero cells.The results showed that V protein was successfully expressed and the positive serum was prepared.The recombinant plasmid pcDNA3.1-CDV-V was expressed in Vero cells and mainly expressed in cytoplasm.The results laid a function for functional studies of CDV V protein.