铁蛋白与口蹄疫病毒VP1在大肠杆菌中融合表达及纳米颗粒自组装

口蹄疫是由口蹄疫病毒引起的世界上最重要的畜牧疾病之一,严重影响世界畜牧业的发展,而疫苗免疫仍然是对疫情预防和控制的最有效手段。铁蛋白具有自组装和生物修饰的特性,在纳米疫苗等领域具有广阔的应用前景。选用O型口蹄疫病毒的vp1基因和幽门螺杆菌铁蛋白基因,通过融合PCR将vp1基因构建到铁蛋白亚基基因前端,在大肠杆菌中表达后通过His标签进行镍柱亲和层析纯化。将纯化好的重组蛋白进行Western blotting检测、质谱分析和透射电镜观察,发现重组蛋白VP1-Ferritin在大肠杆菌中获得表达,并可自组装成纳米颗粒。

Fusion Expression of Ferritin and Foot-and-Mouth Disease Virus VP1 in Escherichia coli and Self-assembly of Nanoparticles

Foot-and-mouth disease is one of the most important livestock diseases in the world caused by foot-and-mouth disease virus,which seriously affects the development of animal husbandry in the world,and vaccine immunization is still the most effective means for the prevention and control of the epidemic.Ferritin has the characteristics of self-assembly and biological modification,and has broad application prospects in the field of nano vaccines.In this study,the vp1 gene of type O foot-and-mouth disease virus and the Helicobacter pylori ferritin gene were selected for nanoparticles assembly.The vp1 gene was fused to ferritin subunit gene by overlap PCR.After being expressed in Escherichia coli,it was purified by nickel column affinity chromatography by His Tag.The purified recombinant protein was then subjected to Western blotting detection,mass spectrometry analysis and transmission electron microscopy observations.It was found that the recombinant protein VP1-Ferritin was expressed in E.coli and self-assembled into nanoparticles.