A Strategy for Searching Antigenic Regions in the SARS-CoV Spike Protein-中国期刊网

摘要 Inthefaceoftheworldwidethreatofsevereacuterespiratorysyndrome(SARS)tohumanlife,someofthemosturgentchallengesaretodevelopfastandaccurateanalyticalmethodsforearlydiagnosisofthisdiseaseaswellastocreateasafeanti-viralvaccineforprevention.Totheseends,weinvestigatedtheantigenicityofthespikeprotein(Sprotein),amajorstructuralproteinintheSARS-coronavirus(SARS-CoV).BaseduponthetheoreticalanalysisforhydrophobicityoftheSprotein,18peptidesweresynthesized.UsingEnzyme-LinkedImmunosorbentAssay(ELISA),thesepeptideswerescreenedintheserafromSARSpatients.Accordingtotheseresults,twofragmentsoftheSgenewereamplifiedbyPCRandclonedintopET-32a.BothSfragmentswereexpressedintheBL-21strainandfurtherpurifiedwithanaffinitychromatography.TheserecombinantSfragmentswereconfirmedtohavepositivecross-reactionswithSARSsera,eitherbyWesternblotorbyELISA.OurresultsdemonstratedthatthepotentialepitoperegionswerelocatedatCodons469-882intheSprotein,andoneepitopesitewaslocatedatCodons599-620.IdentificationofantigenicregionsintheSARS-CoVSproteinmaybeimportantforthefunctionalstudiesofthisvirusorthedevelopmentofclinicaldiagnosis.

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A Strategy for Searching Antigenic Regions in the SARS-CoV Spike Protein

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A Strategy for Searching Antigenic Regions in the SARS-CoV Spike Protein

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