Abstract:
The gene encoding human EC SOD mature peptides ( EC SOD cDNA) was inserted into E.coli expression plasmid pET 28a(+) which contained the T7 promotor, then it was transformed into E.coli BL21(DE3). After induced with 1mmol/L IPTG, the recombinant human EC SOD was highly expressed as inclusion body. SDS PAGE analysis revealed that recombinant EC SOD was cumulated up to 26% of total soluble protein of E.coli cells. After purifying with Ni 2+ IDA Sepharose 6B column, denaturing and refolding, the specific enzymatic activity of recombinant human EC SOD was 1 200U every 1mg purified products. Also, the EC SOD cDNA was inserted into the donor plasmid pFastBacHTb. After transposition, transfection and amplification, the recombinant bacularviruses were infected Tn cells where EC SOD was highly expressed. SDS PAGE and Western blot analysis revealed that the molecular weight of expression human EC SOD was 28 ku, its specific activity was 260U every 1mg Tn cell lysates by pyrogallol autoxidation assay.