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1、VIRUSGENES4:4,289-301,19909KluwerAcademicPublishers,ManufacturedinTheNetherlandsSequenceAnalysisoftheLeaderRNAofTwoPorcineCoronaviruses:TransmissibleGastroenteritisVirusandPorcineRespiratoryCoronavirusKEVINW.PAGE,1PAULBRITTON,1ANDMICHAELE.G.BOURSNELL2A.F.R.C.,InstituteforAnim
2、alHealth,IComptonLaboratory,Compton,Newbury,Berkshire;2HoughtonLaboratory,Houghton,Huntingdon,CambridgeshireReceivedJanuary18,1990AcceptedMarch14,1990RequestsforreprintsshouldbeaddressedtoPaulBritton,A.F.R.C.,InstituteforAnimalHealth,ComptonLaboratory,Compton,Newbury,Berkshire
3、RG16ONN,UnitedKingdomKeywords:transmissiblegastroenteritisvirus,porcinerespiratorycoronavirus,leadersequence,primerextensionAbstractTheleaderRNAsequencewasdeterminedfortwopigcoronaviruses,tranmissi-blegastroenteritisvirus(TGEV),andporcinerespiratorycoronavirus(PRCV).Primerexte
4、nsion,ofasyntheticoligonucleotidecomplementarytothe5'endofthenucleoproteingeneofTGEVwasusedtoproduceasingle-strandedDNAcopyoftheleaderRNAfromthenucleoproteinmRNAspeciesfromTGEVandPRCV,thesequencesofwhichweredeterminedbyMaxamandGilbertcleavage.Northernblotanalysis,usingasynthet
5、icoligonucleotidecomplementarytotheleaderRNA,showedthattheleaderRNAsequencewaspresentonallofthesubgenomicmRNAspecies.TheporcinecoronavirusleaderRNAsequenceswerecomparedtoeachotherandtopublishedcoronavirusleaderRNAse-Thenucleotidesequencedatareportedinthispaperhavebeensubmitted
6、totheEMBL/Genbank/DDBJnucleotidesequencedatabasesandhavebeenassignedtheaccessionnumbersX52157,X52668.290PAGE,BRITTONANDBOURSNELLquences.Sequencehomologiesandsecondarystructuresimilaritieswereidenti-fiedthatmayplayaroleinthebiologicalfunctionoftheseRNAsequences.IntroductionTran
7、smissiblegastroenteritisvirus(TGEV)andporcinerespiratorycoronavirus(PRCV)belongtothefamilyCoronaviridae,alargegroupofpleomorphicenvel-opedviruseswithapositive-strandedRNAgenome.TGEVcausesgastroenteritisinpigs,resultinginahighmortalityinneonates(1).PRCVwasisolatedinseveralEurop
8、eancountriesbetween1984and1986(2-4),doesnotcausediarrhea,andh
