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1、JournalofGeneralVirology(1992),73,3289-3294.PrintedinGreatBritain3289Bovineeoronavirusspikeglycoprotein:localizationofanimmunodominantregionattheamino-terminalendof$2Jean-FrancoisVautherot,1.J.LaporteIandPascalBoireau21LN.R.A.UnitbdeVirologieetImmunologieMolbculaires,C.R.J.J.-DomainedeVilvert,
2、F78352Jouy-en-JosasCedexand2C.N.E.V.A.LaboratoireCentraldeRecherchesV~tkrinaires,22rueP.Curie,F94700Maisons-Alfort,FranceWehaveidentifiedthebindingsiteofmonoclonalTyr798).MAbsreactedmainlywithsixconsecutiveantibodies(MAbs)againstthe$2subunitofthebovineoverlappingpeptideswiththesequenceTTGYRFTN
3、-coronavirusspike(S)glycoprotein.ThelocationofthisFEPFTV.Polyclonalantibodiesfromhyperimmu-sitewasfirstinvestigatedbyusingprokaryoticex-nizedorconvalescentanimalsreactedonlywiththepressionofDNArestrictionfragmentscoveringtherecombinantproteinsidentifiedbyMAbs,andtheentireSgene.Theaminoacidsequ
4、encecontainingthehyperimmuneserumboundtothesamesetofpeptides.antibodybindingsitewasshortenedfrom70to20Thissuggeststhatthishighlyconservedlinearanti-aminoacidsbydigestionofplasmidDNAwithexo-genicdeterminantcorrespondstoanimmunodomi-nucleaseIII,followedbysequencingofthesmallestnantregion.Thisreg
5、ionresemblesbothinlocationanddigestionproductencodinganimmunoreactivefusionimmunodominancethelineardeterminantdefinedonprotein.Finallywesynthesizedasetofnonapeptidestheinfectiousbronchitisvirus$2subunit.Thepresencecoveringthe20aminoacidsequenceextendingfromofsimilarregionsintheN-terminalregion
6、ofthe$2theN-terminalresidueofthe$2subunit(Ala769tosubunitofothercoronavirusesisdiscussed.Bovinecoronavirus(BCV),anentericcoronavirus,etal.,1991;Vautherotetal.,1992).The$2subunithascausesacuteenteritisinnewborncalvesandchronicbeenpredictedtocontaintwolongaheliceswhichareinfectionsinadultcattle(
7、Stairetal.,1972;Crouchetal.,involvedintheformationofthestalkofthepeplomer1985).Whenexaminedbyelectronmicroscopy,thevirus(DeGrootetal.,1987;Boireauetal.,1990),andelicitsdisplaystwofringesofspikes(King&Brian,1982;Deatheproductionofgroup-s
