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1、JournalofGeneralVirology(1990),71,271279.PrintedinGreatBritain271LocalizationofantigenicsitesoftheE2glycoproteinoftransmissiblegastroenteritiscoronavirusIsabelCorrea,1FhtimaGebauer,1MariaJ.Builido,1CarlosSufie,1MarcF.D.Baay,2KornelisjeA.Zwaagstra,2WillemP.A.Posthumus,3JohannesA.Lenstra2
2、andLuisEnjuanes1.1CentrodeBiologiaMolecular,CSIC-UAM,FacultaddeCiencias,UniversidadAut6noma,CantoBlaneo,28049Madrid,Spain,ZlnstituteofInfectiousDiseasesandImmunology,VeterinaryFaculty,StateUniversity,P.O.Box80.165,3508TDUtrechtand3CentralVeterinaryInstitute,P.O.Box65,8200ABLelystad,TheN
3、etherlandsFourantigenicsitesoftheE2glycoproteinoftransmis-productcontainingresidues1to325.ThesamesiblegastroenteritisvirusweredefinedbycompetitiveexpressionproductwasrecognizedbysiteC-specificradioimmunoassaysofmonoclonalantibodies(MAbs).MAbs.ThesedataindicatethatresidueswithintheHere,w
4、edescribethelocalizationofthesesitesbysequence1to325contributetositeCandpossiblyalsotestingtheantigenicityofproteinfragmentsandtositeA.SequencingofmarmutantsthatescapedprokaryoticexpressionproductsofE2genefragments,neutralizationbysiteA-specificMAbsindicatedthatandbysequencingofMAb-resi
5、stant(mar)mutants.residues538and543alsobelongtositeA.ThebindingPartialproteolysisofpurifiedE2proteinallowedtheofsite-specificMAbstoexpressionproductsledisolationofa28KfragmentrecognizedbybothsitedirectlytothelocalizationofsitesBandD,betweenA-andsiteC-specificMAbs.Anantiserumagainstresid
6、ues1to325and379to529,respectively.Thefirstthisfragmentboundtoasyntheticpeptidecontain-37%ofthepolypeptidechainofE2appearstobemoreingresidues1to18andtoanexpressionimmunogenicthantherestofthesequence.IntroductionN-terminal28Kpeptidecouldbeisolatedwhichcon-tainedtheantigenicsiteCandpossibl
7、yalsopartofsiteTransmissiblegastroenteritis(TGE)virusisacorona-A.Theuseofrecombinantantigensledtolocalizationofvirusthatcausesentericdiseaseinswineofallages.ThesitesBandCwithinresidues1to325andofsiteDwithindiseaseisespeciallysevereinnewbornanimalslessthanresidues379to529.Theseq
