人锰超氧化物歧化酶基因的克隆及高表达  

Cloning and Expression of Manganese Superoxide Dismutase Gene in E.coli

作　　者：陶开华[1] 张锦海[1] 李越希[1] 

机构地区：[1]南京军区军事医学研究所,南京210002

出　　处：《药物生物技术》2002年第6期313-316,共4页Pharmaceutical Biotechnology

摘　　要：为获得高表达人锰超氧化物歧化酶(MnSOD)的工程菌。从肿瘤组织中提取总RNA ,利用逆转录聚合酶链反应扩增人MnSOD的cDNA ,将克隆的基因片段插入表达载体pET2 8a(+ )内的NcoI/EcoRI位点 ,转化大肠杆菌BL2 1(DE3 ) ,用PCR及SDS -PAGE的方法筛选高表达人MnSOD的工程菌。结果构建成功高表达人MnSOD的工程菌 ,表达的人MnSOD相对分子质量约为 2 2 0 0 0 ,表达量约占菌体蛋白的 3 0 %。为大量制备人MnSOD和进一步的应用研究奠定了基础。To obtain the engineering E.coli for high expression of human manganese superoxide dismutase(MnSOD) gene, the cDNA of human MnSOD was cloned by RT-PCR technique using the extracted total RNA of human tumor as templates. The cDNA of MnSOD was cloned into NcoI/EcoRI sites in plasmid pET28a(+).This recombinant plasmid was transformed into E.coli BL21(DE3), and screened the engineering E.coli expressing highly MnSOD by PCR and SDS-PAGE. The engineering E.coli expressing MnSOD was established. The study lay a good basis of large scale purification and further practical research of human MnSOD.

关 键 词：锰超氧化物歧化酶 基因克隆 基因表达 

分 类 号：Q786[生物学—分子生物学] Q55