O型口蹄疫病毒VP0和VP1蛋白的可溶性表达与反应原性分析

为了高效可溶性表达O型口蹄疫病毒(FMDV)VP0、VP1结构蛋白,根据大肠杆菌密码子的偏爱性优化合成VP0和VP1基因片段,并将其克隆到p E-SUMO载体中,构建重组质粒SUMOVP0和SUMO-VP1,将重组质粒转化到大肠杆菌BL21(DE3)感受态细胞中进行诱导表达,并优化诱导温度、时间和IPTG浓度等表达条件。结果显示,SUMO-VP0可溶性蛋白表达的最佳条件为:20℃条件下,0.1 mmol/L IPTG诱导表达8 h;SUMO-VP1可溶性蛋白表达的最佳条件为:37℃条件下,0.1 mmol/L IPTG诱导表达12 h。SDS-PAGE电泳和Western blot结果表明,表达的SUMOVP0、SUMO-VP1可溶性蛋白能够被抗FMDV的阳性血清识别,具有很好的反应原性。

Soluble Expression and Immuneoreactivity Analysis of FMDV VP0 and VP1 Protein

To obtain efficient soluble expressive VP0 and VP1 protein of O-type foot-and-mouth disease virus ( FMDV) . We optimized VP0 and VP1 gene according to the preference codon usage of E. Coli. The FMDV structural protein VP0 and VP1 genes were synthesized and cloned into pE-SUMO vector. These two recombinant plasmids,SUMO-VP0 and SUMO-VP1 were transformed into E. coli BL21(DE3) compe-tent cells. Through optimizing the inducing temperature,time and the concentration of IPTG,we got that the optimized expression conditions of SUMO-VP1 was induced by 0. 1 mmol/L IPTG,and induced at 20 ℃ for 8 h . The best inducing conditions of SUMO-VP0 was induced by 0. 1 mmol/L IPTG,and ex-pressed 12 h at 37℃. SDS-PAGE and Western blot analysis showed that the soluble protein could be identified by standard positive serum of FMDV,which confirmed that the fusion protein had good responsiveness.