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1、JournalofGeneralVirology(1991),72,579-587.PrintedinGreatBritain579Sequencecomparisonofthe5'endofmRNA3fromtransmissiblegastroenteritisvirusandporcinerespiratorycoronavirusKevinW.Page,tKarenL.MawdittandPaulBritton*AFRCInstituteforAnimalHealth,ComptonLa
2、boratory,Compton,Newbury,BerkshireRG16ONN,U.K.AnalysisofporcinetransmissiblegastroenteritisvirusORF-3ainTGEV,wasabsentfromthePRCVgenome(TGEV)andporcinerespiratorycoronavirus(PRCV)butapotentialsitewasfoundinthePRCVgenomemRNAspeciesindicatedadeletionin
3、mRNA3ofupstreamofageneequivalenttoTGEVORF-3b.PCRPRCV.Polymerasechainreaction(PCR)wasusedtoanalysis,usingprimerscorrespondingtosequencesclonethe5'endofmRNA3fromPRCVforwithintheORF-3bgeneandtheleaderRNAsequence,comparisonwiththeequivalentregioninTGEV.S
4、mallconfirmedthattheleaderRNAbindingsitewasdeletionswereobservedwithinandaroundthePRCVupstreamofageneequivalenttoTGEVORF-3bonsequenceequivalenttotheputativeopenreadingframePRCVmRNA3butupstreamofORF-3aonTGEV(ORF)ORF-3aidentifiedinTGEV.ThepotentialmRNA
5、3.ThepresenceofthenewleaderRNAbindingRNApolymerase-leadercomplexbindingsite(leadersitewouldberesponsibleforgeneratingthesmallerRNAbindingsite),ACTAAAC,foundupstreamofmRNA3speciesfoundinPRCV-infectedcells.IntroductionpigstoinfectionwithPRCVcouldnotbed
6、istinguishedfromTGEVbyavailablediagnostictests.Transmissiblegastroenteritisvirus(TGEV),firstisolat-TGEVandPRCVvirionscontainthreemajoredbyDoyle&Hutchings(1946),causesgastroenteritisinstructuralpolypeptides;asurfaceglycoprotein[spike(S)]pigsresultingi
7、nhighmortalityandmorbidityinwithamonomericMrof200000,aglycosylatedneonates.Thevirusreplicatesinanddestroysthemembraneprotein(M),observedasaseriesofpolypep-enterocytescoveringthetipsofthevilliinthesmalltidesbetweenM~28000and31000andabasicintestine,res
8、ultingindiarrhoeaanddehydration.TGEVphosphorylatedprotein[thenucleoprotein(N)]ofMrreplicationhasalsobeenshowninlungtissuebyimmune47000associatedwiththeviralgenomicRNA(Garwesfluorescenceandvirusisolation(Underdahletal.,1974).&Pocock,1975;Brittonetal.,
