融合表达的副溶血弧菌不耐热性溶血毒素包涵体复性的研究

目的 构建融合表达载体，获得具有生物活性的副溶血弧菌不耐热性溶血毒素。方法 克隆tlh基因，构建表达载体pET32a ／tlh，融合表达副溶血弧菌不耐热性溶血毒素，用8mol／L的尿素溶解包涵体．亲和层析纯化目的蛋白．采取逐步透析、降低蛋白浓度、加人氧化还原剂等复性方法。结果 复性蛋白具有溶血活性及免疫原性。结论 通过克隆tlh基因，构建融合表达载体pET32a -tlh，采取纯化、复性方法，获得具有融血活性及免疫原性的副溶血弧菌不耐热性溶血毒素。

Renaturation of thermolabile hemolysin inclusion bodies expressed by Vibrio parahaemolyticus]

OBJECTIVE: To clone and construct expression vector pET32a+-tlh to acquire biologically functional thermolabile hemolysin of Vibrio parahaemolyticus. METHODS: tlh gene was cloned and the expression vector pET32a+-tlh constructed. The tlh gene of Vibrio parahaemolyticus was expressed in DE3 in the form of inclusion body, which was resolved in 8 mol/L urea followed by purification of the fusion protein using affinity chromatography and renaturation through gradient dialysis, protein concentration reduction and oxidoreduction. RESULTS AND CONCLUSION: The purified and renatured protein possessed hemolytic and immunogenic activities.