RESUMO
Objective:To highly express HIV-1p17 in E.coli and purify the protein.Methods:①Recombinant plasmid was constructed by inserting HIV-1p17 gene amplified by PCR into plasmid vector,pET28c;②The recombinant plasmid was expressed in BL21,BL21(DE3),BL21(DE3) plysS and HMS174(DE3) of E.coli separately;③The target protein were purified with Ni-NTA resin;④The purified protein was detected by western blot and ELISA.Results:The expression of the P17 protein in BL21(DE3) represented up to 32% of total protein in E.coli,which was the most amounts compared with other kinds of E.coli.The purity of the purified protein reached 95%.The purified protein was recognized by HIV-1P17McAb as well as by HIV-1 positive serum.Conclusion:The recombinant plasmid is highly expressed in BL21(DE3) of E.coli that can be proceeded to the immunocompetence and the bioactivity research.The method of Ni-NTA resin is simple with low protein losing and high purity.And the purified p17 can be employed in early detection of HIV-1 infection and prediction of the clinical progression.
