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1、J~gen.Virol.(1989),70,155-164.PrintedinGreatBritain155Keywords:coronavirus,bovine/nueleotidesequence/haemagglutininCloningandinvitroExpressionoftheGenefortheE3HaemagglutininGlycoproteinofBovineCoronavirusByMICHAELD.PARKER,*GRAHAMJ.COX,DIRKDEREGT,tDAVIDR.FITZPATRICKANDLORNEA.B
2、ABIUKVeterinaryInfectiousDiseaseOrganizationandDepartmentofVeterinaryMicrobiology,UniversityofSaskatchewan,124VeterinaryRoad,Saskatoon,SaskatchewanS7N0WO,Canada(Accepted4October1988)SUMMARYAcDNAclonerepresentingthegenefortheE3glycoprotein,thehaemagglutinin,ofbovinecoronavirus
3、wasisolatedfromaplasmidcDNAlibraryoftheviralgenomeandsequenced.Thegeneislocatedimmediately5'oftheE2glycoproteingeneontheviralgenome.NucleotidesequencingoftheE3genepredictsapolypeptideof424aminoacidswithanMrof47K.InvitrotranslationofmRNAtranscribedfromtheclonedE3geneyieldedapo
4、lypeptideofMr45K,similartothatpredictedfromthenucleotidesequence.Inthepresenceofmicrosomalmembranes,theinvitroproductwascotranslationallyprocessedtoa62KpolypeptidewhichcomigratedonSDS-polyacrylamidegelswiththeE3monomer(gp62)obtainedfromvirus-infectedcells.Boththe45Kand62Kpoly
5、peptideswereimmunoprecipitatedwithE3-specificmonoclonalantibodies,confirmingtheidentityofthegeneasthatencodingtheE3glycoprotein.Finally,onlymonoclonalantibodiestotheE3proteininhibitedhaemagglutinationbythevirusthusconfirmingitsidentityasthehaemagglutininofbovinecoronavirus.IN
6、TRODUCTIONBovinecoronavirus(BCV)isamemberofthefamilyCoronaviridae,agroupofenveloped,positive-strandedRNAviruseswhichinfecthumanandanimalhosts,causinganumberofdiversediseases(Siddelletal.,1983).Inneonatalcalves,BCVcausesanacuteandoftenfatalenteritischaracterizedbyintestinalvil
7、lousatrophyanddiarrhoea(Mebus,1978).Coronavirusesmultiplyinthecytoplasmofinfectedcellsandproducegenomesize(20to27kb)andmultiplesubgenomicmRNAswhichforma3'coterminalnestedset(Siddelletal.,1983;Jacobsetal.,1986;deGrootetal.,1987;Boursnelletal.,1987;Kecketal.,1988;Stern&Kennedy,
8、1980).Ingeneral,onlythe'unique'5'endofeachmRNA(sequencesnotcontained
