猪源戊型肝炎病毒ORF2部分基因的原核表达及其鉴定

摘要

本试验旨在表达猪源戊型肝炎病毒(sHEV) ORF2部分基因片段,纯化表达产物并进行抗原性分析.以sHEV的cDNA为模板,PCR扩增ORF2基因片段,构建重组质粒pET32a-dORF2,转化表达菌BL21,IPTG诱导,SDS-PAGE凝胶电泳和Western blotting进行检测.试验结果表明,获得60 ku的目的蛋白,该蛋白具有良好的抗原性,主要以可溶性形式存在.%The assay was aim to express swine hepatitis E virus (sHEV) ORF2 partial gene fragments, purify the expression product and analyze the antigenicity. ORF2 gene was amplified by PCR from sHEV cDNA as a template,and the recombi-nant plasmid pET32a-dORF2 was constructed, then transformed into the expression bacteria BL21. After IPTG induction, the protein was analyzed by SDS-PAGE and Western blotting. The results showed that the protein was expressed from ORF2 gene in the form of soluble, the protein was 60 ku. The result of Western blotting analysis indicated that the protein could react with the HEV positive serum.