日本乙型脑炎病毒分离株E基因的克隆及序列分析

摘要

[目的]对日本乙型脑炎病毒分离株E基因进行克隆及序列分析.[方法]根据乙型脑炎病毒SA14和SA14-14-2株序列,设计并合成一对E基因的特异性引物,用RT-PCR方法扩增出JEV分离株E基因片段,将其与pMD19-T载体连接,应用DNAStar、Clustal_1.81和Mega序列分析软件进行同源性比较.[结果]分离株E基因的cDNA长1500bp,可编码500个氨基酸；分离株E基因与VN50/Viet Nan/1989/Human brain株、SA14株、Whe株、Benjing1株、P3株、KV1889株、JaGAr01株和SA14-14-2株的核苷酸同源性为88.0％～99.1％,氨基酸同源性为97.8％～99.8％;分离株为乙型脑炎病毒强毒株.[结论]该研究为乙脑病毒基因工程疫苗的研发奠定了一定的基础.%[ Objective] The aim was to clone and analyze the sequence of E genes of Japanese encephalitis virus isolated strain. [ Method] Arnpair of specific primers of E genes were designed according to the gene sequence of JEV SA14 and SA14-14-2 strain. The specific fragment ofrnisolated strain E genes had been successfully amplified hy RT-PCR and sequenced after being cloned directly into the pMD19-T. The homologyrnof nucleotide and deduced amino acid between E genes with different strains which shown on GenBank by the softwares of DNAStar.,rnClustal_1.81 and Mega was compared. [ Result]The results showed that the cDNA of E genes was 1 500 bp, and the homology of 88.0% -rn99.1% in nucleotide and 97. 8% -99. 8% in amino acid with the VN50/Viet Nam/1989/Human brain, SAW, Whe, Benjing 1, P3,rnKV1889, JaGArOl and SA14-14-2 strain. The isolated strain was a virulent strain. [ Conclusion] The study lays a foundation for research andrndevelopment of the genetic engineering vaccine of Japanese encephalitis virus.