中国生物工程杂志

The main reason of Multidrug resistence is P-gp overexpressed in vivio. Because of its expression and localization, it has been suggested that P-gp plays an important role in cancer chemotherapy. In this study, we performed a method for monitoring P-gp overexpressed. A recombinant plasmid named pETP-gp was constructed by cloning MDR 1 1kb gene into prokaryotic expression vector pET-28b(+). E.coli competent host BL21(DE3) and BL21(DE)lyss were transformed with pETP-gp. The recombinant protein P-gp was high level expression in the two Ecoli BL21(DE3) and BL21(DE)lyss after induced by IPTG. Recombinant proteins were used as detecting antigen, and then the optimal reactive condition and the concentration of the second antibodies were confirmed. The expression in BL21(DE3)plyss was higher until to 48% of the total protein of the induced recombinant bacteria. Expression protein was shown as specificity P-gp by immunoprint. It was the first report that MDR 1 1kb gene product was expressed with high level in E.coli. Indirect ELISA for the detection of serum antibodies against MDR 1 was established with the E.coli high expressed recombinant pETP-gp protein as antigen and the rabbit against human IgG by tagging HRP as the second antibodies.