Expression of HIV-1 Gag Protein in Insect cells by Bac-to-Bac System

Objective To express Hiv-1 Gag protein subclass B in insect cells.Methods Recombinant plasmid pfastGag was constructed by inserting the full length of HIV-1 Gag subclass B gene into baculovirus transfer vector pfasbacI.After transforming it into competent E.coli DH10 cells and plate selection,the recombinant shuttle plasmid Bacmid was extracted and transfected to Sf9 cells.In this way,the recombinant baculovirus carrying HIV-1 Gag gene was obtained.The expressed product was confirmed by SDS-PAGE, Western blot and immunofluorescence test. Results The expressed recombinant HIV-1 Gag protein showed good antigenicity. It could be recognized with standard HIV-1 positive serum and anti-HIV-1 p24 McAb. Conclusion The expressed products can be used for the diagnosis of HIV-1 infection and the development of AIDS vaccine.