幽门螺杆菌HspA-UreB融合蛋白包涵体的纯化及活性鉴定

目的：建立一种有效的纯化幽门螺杆菌热休克蛋白A亚单位-尿素酶B亚单位(HspA-UreB)融合蛋白包涵体的方法。 方法：基因工程重组大肠杆菌BL21(pET-HU27)诱导表达的HspA-UreB融合蛋白包涵体经洗涤、变性、复性，在A¨KTA FPLC上运用HiTrap Chelating HP分离纯化，用SDS-PAGE和Bradford法检测目的蛋白的纯度和含量，用Western blotting对纯化后蛋白进行免疫学活性鉴定。 结果：纯化后HspA-UreB融合蛋白的纯度＞90%，含量达0.25 g•L^-1，并可以被HspA-UreB融合蛋白免疫小鼠血清识别。 结论：成功地建立了从包涵体中纯化高纯度HspA-UreB融合蛋白的方法。

Purification and characterization of recombinant HspA-UreB fusion protein of Helicobacter pylori from inclusion body

Objective To establish an effective method for purificating recombinant HspA-UreB fusion protein of Helicobacter pylori from inclusion body. Methods The inclusion bodies of HspA-UreB fusion proteins expressed in recombinant E.coli BL21(pET-HU27) were washed, denatured and renatured. The fusion proteins were isolated and purified by A¨KTA Fast Protein Liquid Chromatogram(FPLC) with HiTrap Chelating HP, then the purity and content of target protein were detected by SDS-PAGE and Bradford method, and the immunocompetence was identified with Western blotting test. Results The recombinant HspA-UreB fusion protein from inclusion body was confirmed to have a purity and content of more than 90% and 0.25 g·L~ -1, respectively, and a high immunocompetence. Conclusion An effective method for purificating recombinant HspA-UreB fusion protein of Helicobacter pylori from inclusion body is established successfully.