HEV-GST融合蛋白的表达、纯化及鉴定

Expression and identification of recombinant HEV-GST fusion protein

目的利用谷胱苷肽转移酶(GST)蛋白纯化系统表达、纯化和鉴定HEV-GST融合蛋白。方法将标记有GST的重组大肠杆菌E.coliBL21诱导表达HEV-GST融合蛋白,采用GST蛋白纯化系统进行纯化、所得产物进行SDS-PAGE和Western Blot鉴定。结果大肠杆菌细胞高效表达出约43Kd蛋白,其分子量与HEV-GST分子量相符,经Western Blot显示所纯化蛋白能被抗GST抗体识别。结论GST大肠杆菌表达系统中高效表达的HEV-GST融合蛋白可用于临床及实验研究。

Objective To express and identify recombinant HEV- GST fusion protein using GST protein purifying system. Methods The recombinant HEV - GST protein was expressed in E. coli cells and the products were purified by GST purifying system. The specific expression was identified by SDS - PAGE and Western ＂blot. Results The HEV - GST fusion protein was highly expressed on 10% SDS - PAGE with molecular weight of 43kD and the fusion protein was identified by anti - HEV antibedy on PVDF membrance. Conclusion The recombinant HEV - GST fusion protein is expressed in E. coli cells efficiently, and the purified HEV- GST can be used in clinical and experimental investgations.