中枢神经系统靶向性Mn-SOD的克隆和在大肠杆菌中的表达研究  

Cloning and Expression of Targeting Mn-SOD to Central Nervous System in E.coli

作　　者：张艳红[1] 贺华君[1] 袁勤生[1] 杨卫东[2] 吴祥甫[2] 

机构地区：[1]华东理工大学生物化学研究所,生物反应器工程国家重点实验室,上海200237 [2]中国科学院上海生物化学与细胞生物学研究所,上海200031

出　　处：《生物化学与生物物理进展》2001年第6期895-899,共5页Progress In Biochemistry and Biophysics

摘　　要：靶向性超氧化物歧化酶 (SOD)是治疗氧自由基引起神经细胞损伤所致疾病等的有效途径 .将破伤风毒素C部分 (tetanustoxinfragmentC ,TTC)基因与编码Mn SOD的cDNA融合克隆进pET 2 2b(+)载体 ,1mmol/L异丙基 D 硫代半乳糖 (IPTG)诱导在大肠杆菌中表达 .SDS 聚丙烯酰胺凝胶电泳 (PAGE)图谱可见约 71ku有表达产物条带 ,与理论计算值相符 ;蛋白质印迹 (Westernblot)分析显示 ,表达产物与抗Mn SOD及破伤风毒素的多抗有免疫反应 ,而且表达产物用邻苯三酚自氧化法测定具有SOD活性 .融合蛋白可作为有效的试剂靶向性输送Mn SOD到神经元细胞 。The nontoxic fragment C of tetanus toxin (TTC) can transport other proteins from the circulation to central nervous system motor neurons. Increased levels of CuZn-SOD are protective in experimental models of stroke and Parkinsons's diseases, where mutations in SOD may cause motor neuron disease. Here the human Mn-SOD is linked to tetanus toxin fragment C gene to construct the fusion gene, then was ligated into prokaryotic expression vector pET-22b (+) expression of the plasmid in E. coli, resulted in the production of a protein has a subunit molecular mass of 71 ku and is recognized by both anti-Mn-SOD and anti-tetanus toxin antibody. The Mn-SOD moiety retains substantial enzymatic activity, where the TTC moiety can delivers the fusion protein to central nervous system neurons. Such fusions should provide a powerful tool for investigating the protective and destructive roles of Mn-SOD in motor neurons.

关 键 词：锰超氧化物歧化酶 破伤风毒C部分 融合表达 中枢神经系统 靶向性运输 人Mn-SOD 克隆 表达 

分 类 号：R394.8[医药卫生—医学遗传学] Q554[医药卫生—基础医学]