Cloning, Sequencing and Expression of
Human Copper,
Zinc-superoxide Dismutase cDNA
SHI Hui-Juan, FAN Li-Qiang, WEI Dong-Zhi and YUAN Qin-Sheng*
( Research Institute of Biochemistry, State Key Laboratory of Bioreactor
Engineering,
East China University of Science and Technology, Shanghai 200237, China
)
SUN Jian-Xing and WU Xiang-Fu
( Shanghai Institute of Biochemistry, the Chinese Academy of Sciences,
Shanghai 200031, China )
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.
Key Words Copper,zinc-superoxide
dismutase; expression vector; gene cloning; gene expression; E. coli
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