狂犬病病毒糖蛋白原核表达及纯化

试验旨在提高狂犬病病毒(rabies virus,RV)糖蛋白(G)在大肠杆菌(E.coli)BL21(DE3)中的表达量。通过优化蛋白质诱导表达的温度、时间、诱导剂浓度等条件,以提高该蛋白质的表达量。SDS-PAGE电泳分析结果显示,重组质粒在1 mmol/mL IPTG、30 ℃诱导6 h条件下蛋白表达量最高,经GST-resin亲和层析柱法纯化获得的纯化蛋白约为36 ku,与预期大小相符。Western blotting结果显示,表达蛋白具有很好的免疫原性和特异性。结果表明,RV G融合蛋白的优化表达和纯化为RV亚单位疫苗及中和抗体的研制奠定了基础。

Prokaryotic Expression and Purification of Glycoprotein Gene of Rabies Virus

In order to improve the expression level of glycoprotein gene of rabies virus (RV) in Escherichia coli BL21(DE3), the optimal conditions for expression of RV G fusion protein (GST-RVG) were analyzed by SDS-PAGE, including different temperatures, different inducing times and concentrations of IPTG. The protein was purified by GST-resin affinity column. The results showed that the expression level of fusion protein expressed in Escherichia coli BL21(DE3) at 30 ℃ for 6 h, and induced with 1 mmol/mL IPTG was highest.The fusion protein was about 36 ku,which was consistent with the expected. The result of Western blotting showed that the fusion protein had good immunogenicities and specificities. Optimizing the expression and purification of RV G fusion protein was benefit for developing sub-unit vaccine and neutralizing antibody of RV.