【摘要】 利用PCR技术扩增马疱疹病毒1型(EHV-1)gD基因主要中和抗原编码区,将其克隆到原核表达载体pGEX-6p-1中,获得的重组质粒pGEX-EHV-gD用IPTG诱导表达。SDS-PAGE电泳分析显示,重组gD蛋白以不溶性包涵体形式表达,与预期大小(43 ku)相符;Western-blotting分析表明,表达产物均能与抗EHV-1和EHV-4阳性血清发生反应。用重组蛋白免疫家兔获得的超免疫血清能中和EHV-1和EHV-4,且中和抗体效价较高,说明原核表达产物具有良好的免疫原性,可以用作ELISA包被抗原或重组基因疫苗。更多还原

【Abstract】 The equine herpesvirus 1(EHV-1) gD gene encoding major neutralizing antigenic domain was amplified by PCR assay and cloned into prokaryotic expression vector pGEX-6p-1.E.coli transformed by the recombinant plasmid pGEX-EHV-gD harboring truncated EHV-1 gD gene was induced by IPTG.SDS-PAGE analysis revealed a fusion protein of 43ku which was expressed in the form of insoluble inclusive bodies.The shared antigenicity of the recombinant protein was confirmed by Western-blotting with antisera against EHV-1 and EHV-4.Hyperimmune sera generated in rabbits immunized with the expressed product could neutralize EHV-1 and EHV-4 with high titers,which indicated its good immunogenicity.The expressed product could be used as potential coating antigen for ELISA or as potential recombinant subunit vaccine.更多还原