鸡补体C3d与禽流感病毒M2融合蛋白的原核表达与反应原性分析

Prokaryotic expression and antigenicity analysis of fusion protein of chicken complement C3d and avian influenza virus M2

为表达禽流感病毒M2蛋白与单拷贝及双拷贝鸡补体C3d的融合蛋白,本实验分别将单拷贝和双拷贝的鸡补体因子C3d基因同禽流感病毒M2基因5'端连接,再将串联基因定向克隆到pET-32a表达载体中进行诱导表达,表达产物用SDS-PAGE和western blot分析。结果表明,表达的2个融合蛋白大小分别约为64ku和97ku,以包涵体形式存在。Western blot分析结果显示,2种重组蛋白可被M2多克隆抗体识别,表明2种重组蛋白具有反应抗原性,为研制以鸡补体C3d为分子佐剂的禽流感新型疫苗奠定了基础。

The M2 protein of avian influenza virus （A/V） is a cross-protectant structure protein, but it is difficult to induce high level immune response because of its poor antigenicity. In this study, one or two copies of C3d gene were ligated with A/V M2 gene and cloned into the expression plasmid vector pET-32（a）, and the expression products were analyzed by SDS-PAGE and western blot. The results showed that the two fusion proteins were about 64 ku and 97 ku and expressed in a form of inclusion bodies. Western blot test showed that the two proteins could react to M2 polyclonal antibody, indicating that the recombinant proteins possessed the antigenicity of M protein. This study laid foundation for research on novel vaccine against AIV by using chicken complement C3d as a molecular adjuvant to enhance the immunogenicity of M2.