Mn-SOD与抗癌胚抗原单链抗体基因的融合及高效表达  

Fusion and Expression of the Gene Encoding Human Mn-SOD to Anti-CEA Single-chain antibody in Escherichia coli

作　　者：贺华君[1,2] 杨卫东[3] 常雅宁 施惠娟[1] 杨冠珍[3] 吴祥甫[3] 袁勤生[1] 

机构地区：[1]华东理工大学生物化学研究所 [2]中国科学院上海生物化学研究所上海200031 [3]中国科学院上海生物化学研究所

出　　处：《生物工程学报》2000年第5期566-569,共4页Chinese Journal of Biotechnology

摘　　要：将Mn SOD与抗癌胚抗原 (CEA)单链抗体基因 (ScFvgene)融合 ,重组到含T7启动子的表达载体 pET 2 2b(+)中 ,构建表达质粒pETMn SOD ScFv ,并转化大肠杆菌BL2 1(DE3) ,进行高效表达 ,表达物占菌体可溶性总蛋白的 2 4%。SDS PAGE和蛋白质印迹图谱显示表达物分子量为 45kD与融合基因编码蛋白质的理论值相符。该蛋白质在大肠杆菌中为分泌型表达有利于纯化。RIA测定表明表达产物能特异性的与抗原CEA结合 ,同时邻苯三酚法测定也表明表达产物具有SOD酶的活性 ,该融合蛋白为分泌CEA肿瘤的靶向性治疗提供新的途径。The gene encoding human manganese\|superoxide dismutase (Mn\|SOD) was fused to anti\|carcinoembryonic antigen single\|chain antibody gene to construct the fusion gene,then was ligated into prokaryotic expression vector pET\|22b(+),The fusion gene was expressed in E.coli at high level,accounting for 24% of the total bacteria soluble protein;and was characterized by SDS\|PAGE and Western\|blot analysis;the expression product had the CEA\|binding ability in RIA,and also had the SOD activity by pyrogallol autoxidation assay.So,the Mn\|SOD moiety retains substantial enzymatic activity,where the ScFv moiety can deliver the fusion protein to tumor,Mn\|SOD is a potential tumor\|suppressor gene,maybe the fusion protein can provide a new pathway to tumor therapy.

关 键 词：癌胚抗原 单链抗体 MN-SOD 融合表达 肿瘤治疗 

分 类 号：Q786[生物学—分子生物学]