Antigen Gene Cloning and Expression of HIV-1 for AIDS Vaccine Design——Ⅲ HIV-1 Antigen Gene Cloning, in Vitro Expression and Antibody Induction

(整期优先)网络出版时间:2002-03-13
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Objective:ToevaluatethehumoralimmuneinductioninratsofacandidateAIDSvaccineexpressingthegagp24genefromasubtypeBHIV-1isolate.Methods:Theamplifiedp24genewasinsertedintoaneukaryoticexpressionvectortoformthesupercoiledDNAvaccine.ThelinearizedexpressedDNAvaccinewaspreparedfromtheexpressionplasmidbypolymerasechainreaction(PCR).TheantigengeneexpressioninratsofthelinearizedandsupercoiledDNAvaccineswereinvitroandinvivodetected.Results:InvitrotranscriptionandNorthernhybridizationshowedthatthelinearizedDNAvaccinecouldsynthesizeamountsofp24mRNAsimilartothesupercoiledDNAvaccine.AntibodyassaysofinoculatedratsconfirmedthatthelinearizedexpressionDNAcouldinduceaslightlyhigherantibodytiterthantheexpressionplasmid,whilethehighestantibodytiterhadbeeninducedbyplasmidplusadjuvantinoculation.Conclusion:TheconstructionofacandidateAIDSvaccinebasedonthep24genecouldshedlightonapotentialHIVvaccine,meritingevaluationinarhesusmacaqueSHIV-AIDSmodel.