绿色荧光蛋白α2b-干扰素融合蛋白的表达与活性研究  

Expression and Characterization of GFP-α2b-IFN Fusion Protein

作　　者：田旺[1] 李正赫[1] 杨文博[1] 白钢[1] 

机构地区：[1]南开大学生命科学学院,天津300071

出　　处：《南开大学学报（自然科学版）》2005年第5期26-30,共5页Acta Scientiarum Naturalium Universitatis Nankaiensis

摘　　要：采用基因重组技术,构建原核表达质粒pET 32a(+)/GFP-α2b-IFN,转化大肠杆菌BL 21,以IPTG诱导融合蛋白的表达并用镍金属螯合层析柱进行纯化.SDS-PAGE及免疫印迹结果显示,46.0 kD的融合蛋白在大肠杆菌中得到了正确的表达,并具有α2b-IFN的抗原活性.受体结合实验表明,该融合蛋白能够与W ISH细胞上的干扰素受体特异性结合,并在激发光下呈亮绿色荧光.采用细胞病变法测定分离纯化的融合蛋白的抗病毒活性,比活力为1.0×107IU/m g.表达的GFP-α2b-IFN融合蛋白具有α2b-IFN的特性与荧光活性,为研究α2b-IFN受体功能以及IFN与细胞相互作用的机理奠定了基础.The prokaryotic expression vector pET32a （＋）/GFP-α2b-IFN was constructed and then transformed to E. coli BL21. The expressed fusion protein after induction with IPTG was purified by nickel chelation chromatography. SDS-PAGE and Western blot analysis showed that the 46.0 kD protein encoded by the fusion gene was expressed correctly in E. coll. The receptor binding experiments indicated that the fusion protein with GFP activity could bind especially to WISH cells containing IFN receptors. After assaying on human WISH cells chal- lenged with VSV, the antiviral activity of the purified fusion protein was 1.0× 10^7 IU/mg. The fusion protein we prepared has not only the green fluorescence but also the α2b-IFN activity. The results can be applied to the research on functional mechanism of α2b-IFN receptor and its reactions with ceils.

关 键 词：绿色荧光蛋白 α2b-干扰素 融合蛋白 原核表达 

分 类 号：Q785[生物学—分子生物学] Q786