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Cloning and Expression of Human Manganese-superoxide Dismutase cDNA
Hui-Juan Shi1, Hua-Jun He, Dong-Zhi Wei
1Research Institute of Biochemistry State Key Laboratory of Bioreactor Engineering East China University of Science and Technology Shanghai 200237 China. qsyuan@ecust.edu.cn
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao Acta Biochimica Et Biophysica Sinica
|July 12, 2002
Abstract:
The cDNA encoding human manganese-superoxide dismutase was amplified from the human liver cell (L02) total RNA by RT-PCR and sequenced. The human Mn-SOD cDNA was ligated into expression vector pET-24a(+) under T7 promoter. After 5 h induction with 1 mmol/L IPTG 800 mol/L Mn(2+) human Mn-SOD was highly expressed in E.coli BL21(DE3). The protein product was up to 50% of the bacteria total protein in soluble form and had specific SOD activity.