牛免疫缺陷病毒基质蛋白和衣壳蛋白的可溶性表达

基质蛋白和衣壳蛋白是BIV的主要结构蛋白,在病毒感染及整个复制周期中起重要作用。本文采用 pTXB系统在大肠杆菌中表达出融合状态的牛免疫缺陷病毒 BIV基质蛋白 MA及衣壳蛋白 CA,经几丁质亲合、自剪切纯化后,获得不含融合片段的纯化产物。每克湿菌体MA产量可达毫克级,CA表达量达十毫克级。用原核表达获得的高纯度CA蛋白免疫大白兔获得的抗血清,经Western Blot 分析显示能够与病毒颗粒的 CA蛋白发生特异反应,证实表达产物具有良好的免疫原性和反应原性,可用于制备相应抗体,为研究 BIV相应基因表达变化,进行体外蛋白质相互作用试验提供工具。

Expression of Matrix and Capsid Portein of Bovine Immunodeficiency Virus

The matrix and the capsid are main structural proteins of Bovine immunodeficency virus (BIV) and play important roles in virus life cycle. In order to analyze these two proteins in vitro, we cloned these two genes into the prokaryotic expression vector pTXB and expressed in E. coli BL21(DE3) in a soluble form. After purification, the matrix without fusion peptide was obtained in milligram magnitude. The capsid was also expressed and purified in ten-milligram magnitude. We used the dissolvable BIV CA to immunized rabbit and got antiserum 53 d after first injection. Western blotting analysis showed that the antiserum could specifically recognize the native HIV CA protein. The protein could be used as the antigen to produce the antiserum and used in protein-protein interaction analysis.