猪伪狂犬病病毒gB蛋白表达及免疫原性分析

摘要

In order to study the expression and immunogenicity of porcine pseudorabies virus (PRV) gB protein,the specific primers were designed with the template of PRV preserved in the laboratory, and the 612 bp conserved gene fragments were amplified and sequenced, then it was cloned into the expression vector pET-28a and transformed into E.coli BL21 (DE3), the target protein was obtained after induced expression and purification.Western blotting was performed to analyse its immunogenicity.The results showed that gB protein was 30 ku, which mainly expressed in the form of inclusion body, and the concentration of the protein was 106 μg/mL, with well reactogenicity.13 PRV positive serum and 16 negative serum in the samples were detected using ELISA Kit on sale, using positive serum, the PRV antibody detection method was initially established with the PRV gB protein as antigen package.%本试验旨在研究猪伪狂犬病病毒(pseudorabies virus, PRV) gB蛋白的表达并分析其免疫原性,以PRV病毒液为模板,设计特异性引物,扩增大小为612 bp的保守片段并测序,将其克隆到表达载体pET-28a中,转化表达菌BL21(DE3),经诱导表达、纯化得到目的蛋白,进行Western blotting分析验证并分析免疫原性.结果表明,表达的gB蛋白大小为30 ku,主要以包涵体形式存在,复性后浓度为106 μg/mL,且具有良好的反应原性.应用市售试剂盒检测到样品中含13份PRV阳性血清和16份阴性血清,利用检出的阳性血清,初步可建立以PRV gB蛋白为包被抗原的PRV抗体ELISA检测方法.