鸭肠炎病毒糖蛋白gB-gD主要抗原域的串联表达

Co-expression of the recombinant protein of gB and gD Genes of Duck Enteritis Virus in Escherichia coli and antigenic analysis of the recombin-ant protein

为了研究鸭肠炎病毒gB和gD基因串联表达后的免疫学活性,根据已发表的鸭肠炎病毒糖蛋白gB和gD基因序列设计两对引物,分别扩增了gB和gD基因主要抗原域,将其克隆到表达载体pPRO-EX—HTb中,构建重组表达质粒pHTb—gB-gD。将重组表达质粒pHTb-gB-gD转化至大肠杆菌BL21(DE3)感受态细胞中,经IPTG诱导表达,SDS—PAGE表明:获得串联表达的gB和gD重组蛋白约60.5 Ku,与预期蛋白的分子质量一致,主要以包涵体的形式存在。Western-blot表明:串联表达的gB和gD重组蛋白可与鸭肠炎病毒阳性血清发生特异性反应,具有良好的抗原性。串联表达的gB和gD重组蛋白为下一步建立针对鸭肠炎病毒的诊断方法奠定基础。

To evaluate the immunogenicity of the co-expressed gB and gD recombinant protein of duck enteritis virus in E. coli, the major antigenic domain of gB and gD gene was amplified by PCR using two pairs of primers and cloned into a prokaryotic expres- sive plasmid pPRO-EX-HTb. The recombinant plasmid pHTb-gB-gD was transformed into the E.coli BL21（DE3） and induced by IPTG. SDS-PAGE showed that the recombinant protein was about 60.5ku and in the form of inclusion body. Western blot analysis showed that the purified fusion protein had reacted with DPV positive serum specifically and had good immunogenicity. These results have laida foundation in the diagnosis of duck enteritis virus.