【摘要】 将构建好的单链抗体 2F3表达载体pTMF2F3ScFv转化到大肠杆菌BL2 1(DE3)。先挑选出表达量高的单克隆 ,然后让其在 37℃进行表达 ,并将表达时的培养条件进行优化。实验结果表明 :最佳诱导条件为开始诱导时的菌体密度OD590nm =1.0～ 1.8,所加异丙基 β D 硫代半乳糖苷 (IPTG)的浓度为 0 3～ 0 5mmol L ,诱导时间 7h ,优化后目的蛋白表达量占菌体总蛋白的 2 0 % ,并用发酵罐成功地进行了扩大培养 ,筛选了洗涤包涵体的最佳条件。采用两步法对包涵体复性进行了研究 ,用Westernblotting及ELISA法检测了所表达的单链抗体及其生物活性 ,并成功制备了含硒单链抗体酶更多还原

【Abstract】 The expression vectors of the gene encoding ScFv-2F3 were transformed into \%E.coli BL21\%(DE3).Clones of higher expression were first selected,then were grown in the presence of IPTG at 37℃ to induce its expression.The culture conditions were carefully optimized.It was found that optimal conditions were as follows:the induction was started as OD 590 reached to 1.0～1.8;the concentration of IPTG was 0.3～0.5mmol/L and induction time is 7h.The yield of ScFv-2F3 expressed in the selected clones is about 20% of the total proteins.The optimal culture conditions were successfully applied to fermenter of 50 L.The conditions of washing the inclusion bodies were also optimized.A two-step method was used to renature the inclusion body.The expression product of interest and its biological activities were characterized with Western blotting and ELISA.A novel selenium-containing single-chain abzyme with GPX activity was prepared.更多还原