幽门杆菌Catalase/GST融合蛋白的表达、标签切除及鉴定  

Expression,Tag Cleavage and Identification of Catalase/GST Fusion Protein of Helicobacter pylori

作　　者：姜茵[1] 奚月[1] 李妍[1] 

机构地区：[1]南方医科大学生物技术学院生物治疗研究所,广州510515

出　　处：《生物技术通报》2012年第2期102-106,共5页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31070119)

摘　　要：旨在利用GST融合基因表达系统表达幽门螺杆菌Catalase融合蛋白,并利用凝血酶切除GST标签。将重组质粒Catalase/pGEX-4T-1转化大肠杆菌BL21(DE3)感受态中,用IPTG进行诱导表达,菌体经反复冻融、溶菌酶裂解及超声破菌后,Catalase/GST融合蛋白以部分可溶性的形式表达在上清中。采用谷胱甘肽琼脂糖树脂Glutathione Sepharose 4B对其进行纯化,得到Catalase/GST融合蛋白,再用凝血酶进行GST标签的切除,所得产物进行Western blotting鉴定。高效表达出Catalase/GST融合蛋白的相对分子质量约85 kD,凝血酶成功地切除了GST标签,Western blotting证实Catalase蛋白能被鼠抗Catalase单克隆抗体识别。It was to express Catalase-GST fusion protein by GST gene expression system and cleave GST-tag using thrombin.The recombinant Catalase/pGEX4T-1 plasmid was transformed into E.coli BL21（DE3） compenent cell and induced by IPTG.The bacterial sediment was lysed by repeating freezing and thawing,lysozyme lysis and ultrasonication.Catalase/GST fusion protein was purified by Glutathione Sepharose 4B and cleaved the GST-tag using thrombin.Then Catalase was identified by anti-Catalase monoclonal antibody using Western blotting.Results showed that the fusion Catalase/GST was partly expressed in soluble form with relative molecular mass of 8.5 kD.After cleavage of GST tag,Catalase protein was recognized by mouse anti-Catalase monoclonal antibody by Western blotting.The recombinant Catalase is successfully expressed and purified,which lay a foundation for further study on function of Catalase protein.

关 键 词：幽门螺杆菌 CATALASE 融合表达 GST标签切除 纯化 

分 类 号：R378[医药卫生—病原生物学]