Expression,purification and identification of HIV-1 gag p17+24 recombinant antigen
Objective To express and purify p17+24 recombinant protein of HIV-1 gag gene for re- search and clinical assay.Methods Coding sequences of gag p17+24 of HIV-1 by PCR were cloned in prokary- otic expression vector pTrcHis2A.Recombinant plasmids were transformed into E.coli BL21(DE3) and induced for protein expression by IPTG.Recombinant proteins were purified by Ni-NTA column chromatography and identi- fied by Western blot.Results SDS-PAGE proved that recombinant proteins showed a band with molecular relative mass (M_r) of 51×10~3.The purified protein was recognized by HIV-1 p17 and p24 McAb.The specificity and sensitivity of p17+24 protein were tested by Western blot using 25 HIV-1 Ab positive and 180 negative sera.Con- clusion Recombinant p17+24 antigen showes high specificity and sensitivity and is a potential diagnostic reagent for HIV-Ab.