传染性法氏囊病病毒VP5蛋白跨膜区的敲除及可溶性原核表达研究

目的]构建敲除传染性法氏囊病病毒（IBDV）VP5蛋白跨膜区序列的原核表达载体，并进行目的蛋白的表达、分离和纯化。方法]利用PCR技术分别扩增IBDVVP5基因的胞外片段和胞内片段，然后将2个片段及pET-28b（＋）同时相接，即载体一胞内片段一胞外片段一载体，构建了敲除跨膜区基因片段的VP5重组表达质粒pET—VP5-FC及改进后的pET—VP5-SC。将表达质粒转化BL21（DE3），IPrG诱导后经Ni亲和层析及凝胶过滤层析纯化重组蛋白。结果]得到可溶性表达的IBDVVP5。结论]为进一步研究VP5蛋白的结构与功能奠定了基础。

Knockout of IBDV VP5 Protein Transmembrane Region and the Soluble Prokaryotic Expression Research

Objective] To construct the prokaryotic expression vector of IBDV VP5 protein whose transmembrane region sequence was knocked out.Moreover,the expression,separation and purification of objective protein were carried out.Method] PCR technology was used to respectively amplify the extracellular and intracellular fragments of IBDV VP5 gene.Then,the two fragments were simultaneously linked to pET-28b(+),and it was the vector-intracellular fragment-extracellular fragment-vector.The recombinant expression vecto...