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人铜锌超氧化物歧化酶cDNA的克隆,测序及表达
引用本文:施惠娟,孙建新.人铜锌超氧化物歧化酶cDNA的克隆,测序及表达[J].生物化学与生物物理学报,1999,31(1):16-18.
作者姓名:施惠娟  孙建新
作者单位:[1]华东理工大学生物反应器工程国家重点实验室 [2]中国科学院上海生物化学研究所
摘    要:用逆转录聚合酶链反应(RTPCR),以人胎肝组织总RNA为模板,扩增了人铜锌超氧化物歧化酶(hCu,ZnSOD)的cDNA,并进行序列分析,将该hCu,ZnSODcDNA重组到T7启动子控制下的分泌型表达载体pET22b(+)中,构建表达质粒pETSOD,并转化大肠杆菌BL21(DE3)。SDSPAGE及蛋白质印迹分析表明,经1mmol/L异丙基硫代βD半乳糖苷(IPTG)诱导后,可高效表达一分子量为19kD的蛋白质,与抗人SOD多抗有特异的免疫反应,表达量约为菌体总蛋白质的30%,具有特异性SOD酶活性,酶活力可达1797u/ml培基。

关 键 词:表达载体  基因克隆  基因表达  Cu  Zn-SOD

Cloning, Sequencing and Expression of Human Copper, Zinc superoxide Dismutase cDNA
SHI Hui Juan,FAN Li Qiang,WEI Dong Zhi and YUAN Qin Sheng.Cloning, Sequencing and Expression of Human Copper, Zinc superoxide Dismutase cDNA[J].Acta Biochimica et Biophysica Sinica,1999,31(1):16-18.
Authors:SHI Hui Juan  FAN Li Qiang  WEI Dong Zhi and YUAN Qin Sheng
Institution:SHI Hui Juan,FAN Li Qiang,WEI Dong Zhi and YUAN Qin Sheng *
Abstract:The cDNA encoding the human copper,zinc superoxide dismutase(Cu,Zn SOD) was amplified from the human liver by RT PCR and sequenced. The cloned human Cu,Zn SOD cDNA was ligated into expression vector pET 22b( ) under T7 promotor. After 3 h induction with 1 mmol/L IPTG, human Cu,Zn SOD was highly expressed in E.coli BL21(DE3). The expression product was up to 30% of the total protein of the bacteria in soluble form, which had specific SOD activity.
Keywords:Copper  zinc  superoxide dismutase  expression vector  gene cloning  gene expression  E  coli  
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