乙型脑炎病毒E蛋白Ⅰ、Ⅱ结构域抗原表位鉴定

Antigenic epitopes mapping of domain I,II of Japanese encephalitis virus envelope protein

为鉴定乙型脑炎病毒(JEV)E蛋白I,II结构域抗原表位,本研究设计了一系列针对E蛋白的I、II结构域(1aa～296aa)部分重叠短肽,分别进行GST融合表达,用JEV阳性血清进行western blot和ELISA反应性筛选,获得了5个线性抗原表位分别为:E1(1FNCLGMGNRDFIEGAS16)、E10-4(77TGEAHNEK84)、E11-2(83EKRADSS YVCKQ94)、E19(145GTTTSENHGNYSAQVG160)和E33(257SQEGGLHHALAGAIVV272)。除E10和E19外,其它表位均为第一次通过实验方法确定的。将这5个融合蛋白对小鼠进行免疫,获得了高滴度的针对5个融合短肽的抗血清,研究证明了这些表位具有良好的免疫原性。本研究为乙型脑炎特异性诊断试剂及表位疫苗的开发奠定了基础。

In order to map the antigenic epitopes domain Ⅰ、Ⅱ of the envelope （E） protein of Japanese encephalitis virus （JEV）, a set of partially overlapping fragments spanning the E protein were fused with GST and expressed. The reactivity of the fusion proteins was detected by western blot and ELISA. Five linear antigenic epitopes, E1 （1FNCLGMGNRDFIEGAS^16）, E10-4 （77TGEA HNEK^84）, Ell-2 （83EKRADSSYVCKQ^94）, El9 （145GTTTSENHGNYSAQVG^160） and E33 （257SQEGGLHHALAGAIVV^272） were identified. Immunization of mice with the fusion proteins revealed that all five proteins could elicit short peptide specific antisera. These results provided important basis for study of the structure and function of domain Ⅰ、Ⅱ of the JEV E protein, and development of diagnostic techniques.