乙型脑炎病毒结构蛋白基因的克隆表达与免疫特性分析

Expression and antigenicity analysis of structural protein of Japanese Encephalitis virus

研究分别克隆了乙型脑炎病毒SA14-14-2株的结构蛋白C、PrM/M及E蛋白基因。将3个结构蛋白基因分别插入表达质粒pET30(a)中,成功构建3个原核表达质粒,转化大肠杆菌BL21(DE3)后利用IPGT诱导,SDS-PAGE分析结果表明,3个结构蛋白获得了表达,表达产物大小与预期一致。Western-blot分析结果表明:表达产物中PrM/M和E蛋白具有很好的免疫反应性,可被抗乙型脑炎病毒血清很好地识别;而C蛋白的免疫反应性较差,不能很好地被乙型脑炎病毒阳性血清识别。该结果提示,乙型脑炎病毒结构蛋白中除E蛋白外PrM/M蛋白也很可能具有诊断抗原的应用潜能。

In this study three structural protein gene of Japanese encephalitis virus （JEV） were first cloned. And expression recombinant plasmid were con-structed by inserted three genes into expression vector pET30（a）. After induction with IPTG and analysis by SDS - PAGE, the results demonstrated that all three proteins expressed in expect molecular weight. The result of Western blot indicated that the PrM/M and E protein could be recognized by sera against JEV. But JEV positive sera nearly could not detect the C protein. This study implies that except for E protein the PrM/M protein may be used as immune diagnose antigen.