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

发布时间：2018-06-25 00:10

  本文选题：口蹄疫病毒 + VP、VP结构蛋白　； 参考：《河南农业科学》2017年02期

【摘要】：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。
[Abstract]:In order to express VP0 VP1 structural protein of foot-and-mouth disease virus type O (FMDV), VP0 and VP1 gene fragments were synthesized according to the preference of E. coli codon and cloned into pE-SUMO vector. The recombinant plasmids SUMOVP0 and SUMO-VP1 were constructed. The recombinant plasmids were transformed into E. coli BL21 (DE3) cells for induction and expression, and the expression conditions such as induction temperature, time and IPTG concentration were optimized. The results showed that the optimal conditions for the expression of soluble protein of SUMO-VP0 were 0.1 mmol / L IPTG at 20 鈩，

本文编号：2063615