PIP融合蛋白在大肠杆菌中的表达纯化及其与PRE特异性的结合  

Expression and purification of PIP fusion protein and its interaction with PRE

作　　者：邓庆丽[1] 黄志明[2] 王巍[1] 邵静[1] 

机构地区：[1]中山医科大学孙逸仙纪念医院医学研究中心,广州510120 [2]中山医科大学孙逸仙纪念医院肿瘤科,广州510120

出　　处：《中华微生物学和免疫学杂志》2001年第4期355-358,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金 (396 70 6 5 3);广东省自然科学基金(96 0 14 3);广东省科委;卫生厅乙肝联合攻关资助项目

摘　　要：目的　表达纯化PIP融合蛋白 ,探讨研究PIP与PRE特异性结合的方法。方法　构建GST PIP融合蛋白表达质粒pGENKS PIP ,表达质粒在大肠杆菌BL2 1中诱导表达GST PIP融合蛋白 ,并用亲和层析柱加以纯化。利用这一融合蛋白通过Northwesternblot、UV crosslinking方法研究PIP与PRE的特异性结合。结果　GST PIP融合蛋白表达质粒pGENKS PIP在大肠杆菌BL2 1中成功地诱导表达了可溶性GST PIP融合蛋白 ,纯化后获得了单一的GST PIP融合蛋白。Northwesternblot和UV crosslinking证实PIP与PRE特异性结合。比较两种方法 ,前者较后者特异性高 ,灵敏度低 ,而后者操作简便。结论　表达和获得纯化的PIP融合蛋白 。Objective To obtain purified PIP fusion protein and to test its interaction with PRE. Methods By constructing pGENKS PIP recombinant, GST PIP fusion protein was induced with Isopropyl β D thiogalactoside(IPTG) to express in E.coli BL21 and purified by glutathione resin. The interaction of GST PIP fusion protein with PRE was determinated by Northwestern blot and UV cross linking. Results Soluble GST PIP fusion protein was purified and identified by Western blot. The interaction of GST PIP fusion protein with PRE was proved by Northwestern blot and UV cross linking. Compared with UV cross linking, Northwestern blot showed higher specificity but lower sensitivity. Conclusion GST PIP fusion protein was expressed and purified. PIP interacted with PRE.

关 键 词：PIP融合蛋白 大肠杆菌 表达 纯化 PRE 紫外交联试验 

分 类 号：R346[医药卫生—基础医学]