从包涵体中纯化重组人幽门螺杆菌热休克蛋白A亚单位  

High purification of recombinant Helicobacter pylori heats shock protein A from inclusion body

作　　者：王缚鲲[1] 邹全明[1] 张卫军[1] 田文标[1] 杨王君[1] 郭学青[1] 郭红[1] 

机构地区：[1]第三军医大学医学检验系临床微生物学教研室,重庆400038

出　　处：《第三军医大学学报》2002年第11期1332-1335,共4页Journal of Third Military Medical University

基　　金：国家"九五"重点科技攻关项目 ( 96 90 1 0 1 54 )

摘　　要：目的　建立一种有效的从包涵体中纯化重组人幽门螺杆菌热休克蛋白A亚单位HspA的方法。方法　重组基因工程大肠杆菌发酵后 ,表达的Hp的HspA包涵体经洗涤、变性、复性 ,采用QSepharoseHighPerformance阴离子交换层析和Superdex 75凝胶过滤层析分离纯化。使用SDS PAGE和HPLC检测纯度 ,选用ELISA和动物实验对纯化蛋白的免疫学活性和生物学活性进行鉴定。结果　Hp的HspA包涵体经洗涤和溶解后 ,Hp的HspA的纯度 >60 %。包涵体溶解液经阴离子交换层析和凝胶过滤层析后纯度超过 95 %。Hp的HspA纯品经检测具有良好的免疫学活性和生物学活性。结论　本研究建立的分离纯化方法可从包涵体中获得高纯度的重组HspA蛋白 。Objective To establish an effective method for obtaining recombinant Helicobacter pylori heat shock protein A (HspA of Hp) with biological activities from inclusion bodies of E. coli . Methods The inclusion bodies of HspA expressed in E. coli were washed, denatured and renatured. The two step chromatographic procedure (Q Sepharose High Performance ion exchange and Superdex 75 size exclusion) were used for the purification. The purity of the Hp HspA was detected with SDS PAGE and HPLC. Specific biological activity of the purified Hp HspA was detected with ELISA and animal test. Results Hp HspA was confirmed to have a purity of more than 95%, and a high specific biological activity. Conclusion The established method provides a sound purification effect for Hp HspA.

关 键 词：包涵体 蛋白纯化 重组人幽门螺杆菌 热休克蛋白 

分 类 号：R377[医药卫生—病原生物学] Q78[医药卫生—基础医学]