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1、J.gen.Virol.(1985),66,2253-2258.PrintedinGreatBritain2253Keywords:inJeetiousbronchitis/coronavirusIBV/mRNAD/nucleotidesequence.SequencingofCoronavirusIBVGenomicRNA:ThreeOpenReadingFramesinthe5''Unique'RegionofmRNADByM.E.G.BOURSNELL,*M.M.BINNSANDT.D.K.BROWNHoughtonPoultryR
2、esearchStation,Houghton,Huntingdon,Cambs.PE172DA,U.K.(Accepted2July1985)SUMMARYThenucleotidesequenceofagenomiccDNAclonecorrespondingtothe5'terminaldomainofmRNADoftheBeaudettestrainofinfectiousbronchitisvirus(IBV)hasbeendetermined.Thisregioncontainsthreeopenreadingframeswh
3、ichpredictpolypeptidesofmolecularweights6700(6.7K),7.4Kand12.4K.Thepredicted12-4KpolypeptidehasacodonusageverysimilartothatpredictedfortheproductsoftheIBVnucleocapsid,membraneandspikegenes.Thesequencealsopredictsahydrophobic,potentiallymembrane-anchoring,regionintheNtermi
4、nalhalfofthe12.4Kpolypeptide,andahydrophilicCterminus.Coronavirusesareenvelopedviruseswithasingle-strandedRNAgenomeofpositivepolarity(Siddelletal.,1983;Sturman&Holmes,1983).Thegenomeofinfectiousbronchitisvirus(IBV)isabout20kilobasesinlength(Stern&Kennedy,1980a;Siddelletal
5、.,1983).InIBV-infectedcellssixmajormRNAspeciesareproduced.ThesemRNAs,designatedAtoF,rangeinlengthfromabout2kbtogenomelength,andhavebeenshowntoshareacommon3'terminusandformanoverlappingor'nested'set(Stern&Kennedy,1980a,b)(seeFig.1).Translationstudiesinvitrohavedemonstrated
6、thatmRNAsA,CandEencodethethreemajorviralproteins:thenucleocapsidprotein,themembraneglycoproteinandtheprecursortothespikeorsurfaceprojectionglycoprotein,respectively(Stern&Sefton,1984).SequencingoftheIBVgenomehasshownthatthecodingsequencesforthesepolypeptideslielargelywith
7、inthe'unique'5'terminalregionofeachmRNAspecieswhichisnotpresentinthenextsmallestmRNA(Boursnelletal.,1984,1985;Binnsetal.,1985).However,nospecifictranslationproductshave,todate,beendetectedfrommRNAsBandD(Stern&Sefton,1984;Boursnell&Brown,1984).SequencingstudiesofgenomicRNA
8、intheregionsofthe5'terminaldomainsofmRNAsBandDhavebeencarriedouttodeterminewhetherthesecontainpo
