鸡传染性法式囊病毒VP2原核表达蛋白的纯化和复性  

Purification and Refolding of Infectious Bursal Disease Virus(IBDV) Structural Protein VP2 Expressed in Escherichia coli

作　　者：樊国燕[1] 胡平[1] 

机构地区：[1]郑州牧业工程高等专科学校,河南郑州450011

出　　处：《安徽农业科学》2008年第18期7639-7641,共3页Journal of Anhui Agricultural Sciences

基　　金：河南省重点科技攻关课题(0423011200)

摘　　要：[目的]探讨鸡传染性法式囊病毒(IBDV)VP2原核表达蛋白的纯化和复性。[方法]对原核表达的IBDVVP2蛋白进行可溶性与不可溶性分析,并利用溶菌酶、Triton-100、尿素等试剂进行纯化和复性,对复性前后的蛋白分别与特异性多抗进行Dot-ELISA。[结果]可溶性与不可溶性分析的结果表明,蛋白主要以包涵体形式存在。Dot-ELISA表明,复性后蛋白的反应活性增强了10倍,对复性后的蛋白与IBDV16株单抗进行Dot-ELISA,其中有11株与其发生特异性反应。[结论]复性后的蛋白与鸡传染性法式囊病毒单克隆抗体的反应性明显提高。[Objective] The research aimed to study the purification and refolding of infectious bursal disease（IBDV） structural protein VP2 expressed in Escherichia coli. [Method] IBDV structural protein VP2 expressed in E. coli was analyzed by soluble and insoluble, and was purified and refolded by using lysozyme, Triton-100 and carbemide. And the Dot-ELISA was made between protein before and after refolding and specific polyclonal antibodies. [Resuit] The results showed that protein was produced in the form of insoluble bodies. Then the protein were sublimated and refolded by lysozyme, Triton-100, carbamide, and the activity of the protein was increased 10 time by Dot-ELISA with polyclonal antibodies compared with protein before purification. Refolded protein were assayed with 16 monoclonal antibodies by Dot-ELISA, and 11 monoclonal antibodies were reactivity. [ Conclusion] The reactivity of protein after refolding with IBDV single polyclonal antibody was improved significantly.

关 键 词：鸡传染性法式囊病毒 VP2蛋白 包涵体 纯化 复性 

分 类 号：S855.3[农业科学—临床兽医学]