乙型脑炎病毒E蛋白抗原表位多肽序列鉴定及分析

Identification of an antigenic epitope on envelope glycoprotein of Japanese encephalitis virus

为了对流行性乙型脑炎病毒(JEV)E蛋白抗原表位E19进行核心序列定位,本研究设计了一系列编码相互部分重叠短肽核苷酸序列,原核融合表达后经westernblot分析,确定其抗原表位核心序列为150ENHGNYS156。该表位在乙型脑炎不同基因型的各种病毒株间为高度保守序列;根据JEVE19表位与同一血清群其他黄病毒之间的差异,在同源序列位置设计了系列突变短肽,融合表达后western blot分析的结果表明,其他黄病毒属病毒同源序列不能与抗JEV阳性血清反应。本实验结果表明JEVE蛋白抗原表位E19具有鉴别检测JEV与西尼罗病毒等其它黄病毒的意义。本实验为进一步研究E蛋白结构和功能以及建立乙型脑炎临床鉴别诊断方法奠定了分子生物学基础。

To identify the antigen epitope on envelope glycoprotein （gE） of Japanese encephalitis virus （JEV）, a series partial overlopping oligo DNAs encoding short peptides were synthesized based on gE within 145GTTTSENHGNYSAQVG160 of E19 epitope region by our previous studies. The core amino sequence of 150ENHGNYS156 was identified by western blot with anti-JEV sera to these short peptides expressed in E. coli. The homologous analysis indicated that the amino acid sequences of the epitope were highly conserved among different JEV strains. Furthermore, another series of short peptides were designed at the position of homologous sequence according to the difference flavivirus strains. Western blot result showed that the homologous sequences of flavivirus strains had no cross-reaction with anti-JEV sera. The findings of the present studies would provided basis for further study of JEV gE and development of differential diagnostic techniques for Japanese encephalitis from other flavivirus infection.