鸡传染性法式囊病毒VP2原核表达蛋白的纯化和复性

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。

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

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 carbamide.And the Dot-ELISA was made between protein before and after refolding and specific polyclonal antibodies.Result] 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.