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)

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。

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

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-b...