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

[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.