【摘要】 目的:构建大肠埃希菌外膜蛋白OmpA原核表达载体,优化OmpA蛋白的表达条件。方法:通过分子克隆获得OmpA蛋白的表达菌株;采用正交试验,获得OmpA菌株的最佳表达条件与培养条件。结果:OmpA重组载体双酶切、DNA测序鉴定与预测结果一致,Western Blot分析表明抗体能与OmpA蛋白结合;OmpA菌株最佳诱导表达条件为:IPTG浓度0.1 mmol/L,诱导时菌液OD600值0.8,28℃诱导8h;最佳培养条件为:葡萄糖浓度1%,转速230r/min,装液量50mL。结论:成功克隆OmpA基因,确定OmpA蛋白菌株的最佳表达条件与培养条件。更多还原

【Abstract】 Objective:To construct prokaryotic expression vector for E.coli outer protein OmpA,and optimize the condition for expression of OmpA.Method:The OmpA expression strain was obtained by molecular clone,and the optimal inducing and culture condition was gained by the method of orthogonal experiment.Result:OmpA recombinant vector had been sequenced and restriction digestion identification to confirm the correct construction.Western Blot results showed that the antibody could bind to OmpA protein.The optimal inducing expressing condition of OmpA protein:IPTG concentration,strain OD600 value,inducing time and temperature,which was 0.1mmol/L,0.8,8h and 28℃,respectively; the optimal culture condition:glucose concentration,rotation rate and medium volume,which were 1%,230r/min and 50mL,respectively.Conclusion:Succession of cloning of E.coli OmpA gene,the inducing and culture conditions of recombinant OmpA was preliminarily optimized.更多还原