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。
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 p E-SUMO vector.These two recombinant plasmids,SUMO-VP0 and SUMO-VP1 were transformed into E.coli BL21(DE3) competent 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 at20 ℃ for 8 h.The best inducing conditions of SUMO-VP0 was induced by 0.1 mmol/L IPTG,and expressed 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.
出处
《河南农业科学》
CSCD
北大核心
2017年第2期105-110,共6页
Journal of Henan Agricultural Sciences
基金
国家重点研发计划项目(2016YFD0500704)
河南省基础与前沿技术研究计划项目(152300410238)
河南省科技创新基础前瞻类项目(20141644)
河南省重大科技专项(141100110100)
