摘要
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8 mol/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、加入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。
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.
出处
《第一军医大学学报》
CSCD
北大核心
2004年第9期995-997,共3页
Journal of First Military Medical University
基金
全军"十五"指令性课题(01L050)~~
关键词
副溶血弧菌
不耐热性溶血毒素
包涵体
复性
溶血活性
Vibrio parahaemolyticus
thermolabile hemolysin
inclusion body
renaturation
hemolysis
