【摘要】 以RT-PCR法扩增犊牛前胸腺素α基因(prothymosin-α,ProT-α),与原核表达载体pGEX-4T-1连接生成重组质粒pGEX/ProT-α,再将重组表达质粒转化大肠埃希菌BL21(DE3)。重组菌经IPTG诱导后表达的GST-ProT-α融合蛋白主要存在于细菌裂解液中。SDS-PAGE电泳表明,GST—ProT-α融合蛋白表达量较高,分子量为38 ku;Western-blot和动物细胞试验表明,该产物能与胸腺素α1抗体发生特异性免疫反应,并可显著提高小鼠脾细胞增殖率和NK细胞杀伤活性。更多还原

【Abstract】 ProT-α genes of calf were clones by RT-PCR.Both the ProT-α genes and procaryotic expression vectors(pGEX4T-1) were digested with EcoR I and BamH I,and linked each other to produce recombinant plasmid pGEX/ ProT-α,and converted E.coli BL21(DE3) with it.After induced with IPTG(isopropyl-beta-D-thiogalactopyranoside) the fusion protein of GST-ProT-α expressed by the recombinant E.coli existed in the bacterial lytic solution.SDS-PAGE electrophoresis indicated that the fusion protein of GST-ProT-α was highly expressed in amount with molecular weight at 38 ku.Western-blot and animal experiment indicated that the product could specifically react immuno-reaction with ProT-α I antibody,and remarkably raise the proliferation of mouse spleen cell and NK cell killing activities.更多还原