口蹄疫病毒结构蛋白VP2-3-1的纯化复性与活性检测

Purification, refold and activity assay of FMDV structural proteinVP2-3-1 expressed in Escherichia coli

收集含有口蹄疫病毒结构蛋白VP2 3 1基因的大肠埃希氏菌菌液 ,将其超声破壁 ,用菌体裂解液裂解 ,提取包涵体 ,并用 8mol/L尿素溶解 ,收集上清液 ,用亲和层析法纯化融合蛋白VP2 3 1 ,透析法复性 ,并用间接ELISA检测FMDV阳性血清和阴性血清。结果 ,P/N值大于 2 .1 。

The structural protein VP2-3-1 of foot-and-mouth disease virus was produced in Escherichia coli in the form ofinclusion bodies. After extaction from the E.colicells ,the inclusion bodies were solubilized and denatured in the presence of 8 mol/L urea. The expressed VP2-3-1 fusion protein was (purified) by Ni-NTA His. Bind Resin affinity chromatography and renatured finally bythedialysismethod. The activity of the purified VP2-3-1 was assayed by ELISA with FMDV positive serum and negative (serum). The result revealed that the purified protein VP2-3-1 has certain activities.