减蛋综合症病毒纤维蛋白C-末端的分泌表达及抗原性分析(英文)

Secretary Expression and Antigenicity Analysis of C-terminal of Fibrin Protein on Egg Drop Syndrome Virus

目的:在大肠杆菌中分泌表达重组纤维蛋白的C-末端序列,并检测其抗原性。方法:采用PCR技术扩增了减蛋综合症病毒(EDSV)纤维蛋白C-末端的编码基因,并将其克隆到组成型分泌表达载体pUC18ompAcat上构建pUC18ompA-EDS。将该重组质粒转化大肠杆菌BL21(DE3)菌株构建工程菌,培养工程菌以表达目的蛋白。结果:SDS-PAGE分析表明,纤维蛋白C-末端在大肠杆菌中成功实现了表达,且部分重组蛋白分泌到了周质空间和胞外的培养基中,Ni2+-NTA树脂分离纯化后,Western blotting对其免疫原性的分析表明,重组蛋白可与鸡抗EDSV血清发生特异反应。结论:说明获得的纤维蛋白的C-末端具有明显的抗原性,该研究对于开发预防EDSV的基因工程疫苗的研究具有一定参考作用。

Objective： To express the recombinant C-terminal of fibrin protein in Escherichia coli,and to investigate the antigenicity.Method： The DNA encoding C-terminal of fibrin protein of egg drop syndrome virus（EDSV） was amplified by PCR,and then cloned into constitutive and secretary plasmid pUC18ompAcat to construct pUC18ompA-EDS.The recombinant plasmid was introduced into E.coli BL21（DE3） to construct engineering bacteria.Result： Recombinant protein could be expressed in engineering bacteria,and partial recom-binant protein was secreted into periplasmic space and culture medium.The recombinant C-terminal of fibrin protein purified by Ni2＋-NTA affinity chromatography could react with antiserum of chicken infected with EDSV.Conclusion： The recombinant protein had obvious antigenicity.This study will offer some reference to the production of gene engineering vaccine against EDSV.