EB病毒早期蛋白P54的原核表达及其免疫原性的研究

PCR法获得编码EB病毒早期蛋白P54的基因BMRFl，序列分析后亚克隆入原核表达载体pET30a。表达质粒pET30a-BMRF1在大肠杆菌BL21(DE3)菌株中经IPTG诱导后表达了P54抗原，SDS—PAGE表明其相对分子质量为51000；采用镍离子亲和柱纯化重组蛋白。Western印迹结果表明纯化蛋白免疫BALB／c小鼠后产生了P54特异性抗体。间接免疫荧光表明免疫血清可以识别激活的Raji细胞中表达的P54蛋白。以上结果表明构建了原核表达质粒pET30a-BMRF1并在大肠杆菌细胞中成功表达EB病毒早期蛋白P54，表达蛋白具有很好的抗原性和免疫原性。

Expression of Epstein-Barr virus early antigen P54 in E.coli andits immunogenicity analysis

Amplification was performed with specific pairs of PCR primer to obtain the coding sequence of BMRF1,and then the PCR fragment was subcloned into the pET30a vector after sequencing.The pET30a-BMRF1was con-structed and recombinant P54protein was expressed in E.coli BL21(DE3)induced with IPTG.SDS-PAGE showed that its molecular weight of the expressed protein was about 51kD.The purified protein was obtained by Ni + affinity chromatography and was used to immunize BALB/c mice.Antibodies were achieved by immunized mice and the specificity of antisera was identified by Western blot assay.IFA demonstrated that the P54protein produced in acti-vated Raji cell could be recognized by the antisera.The result showed that recombinant P54protein expressed in E.coli showed high antigenicity and immunogenicity.