鸭病毒性肠炎gB基因的原核表达及产物的抗原性分析

利用DNAStar分析并筛选出鸭病毒性肠炎病毒gB基因的两段主要抗原区域,PCR扩增出这两段主要抗原区域并克隆进入载体pMD18-TVector,目的基因经PCR和酶切鉴定后连接至原核表达载体pET-32a(+)。获得的重组质粒分别转化E.coliBL21(DE3)感受态细胞,SDS-PAGE电泳分析显示重组蛋白在IPTG诱导下以包涵体的形式高效表达,Western-bloting及免疫鼠血清ELISA抗体检测结果表明重组蛋白具有良好的免疫原性。

Prokaryotic expression of duck enteritis virus gB gene and antigenicity analysis of the expressed product

Two fragments of duck enteritis virus gB gene encoding major protective antigen regions were chosen on the basis of DNAStar software and obtained by PCR amplification.Then they were cloned into pMD18-T vector.After confirmed by PCR and restriction endonuclease analysis,the recombinant vectors were constructed using prokaryotic expression vector pET-32a( ).The recombinant plasmids were transformed into competent E.coli BL21(DE3) for further protein expression and the result of SDS-PAGE analysis indicated that the recombinant proteins could be expressed in inclusion body form at a high level in the presence of IPTG.The western-blotting analysis and ELISA showed the good antigenicity of the recombinant proteins.