鸡传染性法氏囊病毒VP2蛋白在大肠杆菌细胞周质中的表达

Expression of VP2 Protein of Infectious Bursal Disease Virus in the Periplasm of Escherichia coli

[目的]利用大肠杆菌可溶性地表达鸡传染性法氏囊病毒(IBDV)VP2蛋白。[方法]根据IBDV vp2全长序列(Gen Bank登入号AY704912),设计一对缺失VP2蛋白N-端80个氨基酸残基的特异性引物,克隆IBDV HQ株的vp2基因,插入质粒p ET-22b中构建大肠杆菌周质表达质粒p ET-22b-vp2,经IPTG诱导后表达重组蛋白。[结果]在SDS-PAGE中可见大小约为39 k Da重组蛋白。重组蛋白纯化后,Western Blot检测表明其具有良好的反应原性。[结论]在大肠杆菌细胞周质中可以高效地可溶性地表达鸡传染性法氏囊病毒IBDV VP2蛋白,并且该重组蛋白有较好的免疫原性,可以为后续亚单位疫苗及检测试剂盒开发提供所需的蛋白。

[Objective] To soluably express the VP2 protein of infectious bursal disease virus by using Escherichia coli system. [Method]Based on the vp2 full-length sequence of IBDV( Gen Bank ID No. AY704912),a pair of specific primers were designed for the amplification of the vp2 sequence which had been deleted the N-terminal 80 amino acid residues of VP2 protein. The truncation gene of VP2 protein of IBDV HQ strain was cloned,and inserted into plasmid p ET-22 b. After IPTG induction,the recombinant protein was expressed in soluble form in the periplasm of E. coli. [Result]The recombinant protein was about 39 k Da as SDS-PAGE shown,and the purified recombinant protein had good immunogenicity as Western Blot shown. [Conclusion] The IBDV VP2 protein could be effectively expressed in the periplasm of E. coli,and the recombinant VP2 protein showed good immunogenicity.