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1、DNAANDCELLBIOLOGYVolume24,Number8,2005©MaryAnnLiebert,Inc.Pp.503–509IdentificationandAntigenicEpitopeMappingofImmunodominantRegionAminoResidues510to672ontheSpikeProteinoftheSevereAcuteRespiratorySyndromeCoronavirusRONG-HONGHUA,YUN-FENGWANG,ZHI-GAOBU,YAN-JUNZHOU,JIN-YINGGE,XI-JUNWANG,andGUANG-
2、ZHITONGABSTRACTThesevereacuterespiratorysyndrome(SARS)isanewlyemerginghumaninfectiousdiseasecausedbythesevereacuterespiratorysyndromecoronavirus(SARS-CoV).Thespike(S)proteinofSARS-CoVisamajorvirionstructuralprotein.Itplaysanimportantroleintheinteractionwithreceptorsandneutralizinganti-bodies.
3、Inthisstudy,theS1domainofthespikeproteinandthreetruncatedfragmentswereexpressedbyfusionwithGSTinapGEX-6p-1vector.Westernblotresultsdemonstratedthatthe510–672fragmentoftheS1domainisalinearepitopedominantregion.Tomaptheantigenicepitopeofthislinearepitopedominantregion,asetof16partiallyoverlappi
4、ngfragmentsspanningthefragmentwerefusedwithGSTandexpressed.FourantigenicepitopesS1C3(539–559),S1C4(548–567),S1C7/8(583–606),andS1C10/11(607–630)wereidentified.Immunizationofmicewitheachofthefourantigenicepitope-fusedproteinsrevealedthatallfourproteinscouldelicitspikeproteinspecificantisera.Al
5、lofthemwereabletobindtothesurfacedomainofthewholespikeproteinexpressedbyrecombinantbaculovirusininsectcells.IdentificationofantigenicepitopesofthespikeproteinofSARS-CoVmayprovidethebasisforthedevelopmentofimmunity-basedprophylac-tic,therapeutic,anddiagnosticclinicaltechniquesforthesevereacute
6、respiratorysyndrome.INTRODUCTIONhomologywithanyotherknownanimalcoronaviruses(Marraetal.,2003;Qinetal.,2003;Rotaetal.,2003).Recently,how-HESEVEREACUTERESPIRATORYSYNDROMEcoronavirusever,SARS-CoV-likeviruseswereisolatedfromHimalayanT(SARS-CoV)isanewlyemergedvirus,andhasbeeniden-palmcivets(Guanet
7、al.,2003),whichhadnotbeenfoundintifiedtobethecausativeagentofthesevererespiratorysyn-anyanimalorhumanbefore2003.Phylogeneticanalysisindi-drome(SARS)(Drostenetal.,2003;Ksiazeketal.,2003;PeiriscatedthatthenewSARS-CoVandSARS-CoV-likevirusesareet
