Cloning, Sequencing and Expression of Human Copper, Zinc-superoxide Dismutase cDNA.
Shi Hui-juan, Liqiang Fan, Dongzhi Wei, Yuan Qin-sheng, Jian-Xing Sun, Xiang‐Fu Wu
Abstract
Shi Hui-juan, Liqiang Fan, Dongzhi Wei, Yuan Qin-sheng, Jian-Xing Sun, Xiang‐Fu Wu
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.
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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.
Key concepts: Complementary DNA, Superoxide dismutase, Cloning (programming), Molecular biology, Zinc, Expression vector, Escherichia coli, lac operon