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1、(c)ELSEVIERAnn.Inst.Pasteur/Virol.Paris19881988,139,123-138SEQUENCEANDANALYSISOFBOVINEENTERITICCORONAVIRUS(F15)GENOMEI.--SEQUENCEOFTHEGENECODINGFORTHENUCLEOCAPSIDPROTEIN;ANALYSISOFTHEPREDICTEDPROTEINC.Cruci6re(2)andJ.Laporte(2)StationdeVirologieetd'Immunologie,INRA,CRJJ,DomainedeVilvert,78350
2、Jouy-en-Josas(France)SUMMARYSequencesencodingtheNproteinofthebovineenteriticcoronavirus-F15strain(BECV-F15)havebeenclonedinPBR322plasmidusingcDNApro-ducedbyprimingwitholigo-dTonpurifiedviralgenomicRNA.Some265insert-containingcloneswerestudied.Hybridizationoftheseinsertswithpo-ly(A)+RNAextract
3、edfrominfectedcellsledtotheconclusionthattheywerelocatedatthe3'-endofthegenome.AftersubcloninginM13phageDNA,clonesweresequencedbytheSangertechnique.A1,710-nucleotidesequencecorrespondingtothegenecodingfortheviralN-proteinwasestablished.Itshows2overlappingopenreadingframes(ORF).The3'-non-codin
4、gendofthegenehasan8-nucleotidesequenceincommonwiththehomologousgenomeareasofMHV,TGEandIBVviruses.ThissequencemayrepresentthepolymeraseRNAbindingsite.AnupstreamsequencesurroundingthefirstAUGofthesmallerORFcor-respondstoapotentiallyfunctionalinitiationcodon.Thesequenceoftheprimarytranslationpro
5、ductdeducedfromtheDNAsequencepredictsapolypeptideof207aminoacids(22.9Kd)withahighleucine(19.8%)con-tent,possessingahydrophobicN-terminalend.ReceivedDecember4,1987.(1)Presentaddress:LaboratoireCentraldeRecherchesV6t6rinaires,22ruePierreCurie,BP67,94703Maisons-AlfortCedex.(2)Towhomallcorrespond
6、enceshouIdbeaddressed.124C.CRUCIEREANDJ.LAPORTEThelargerORFhasacodingcapacityof448aminoacids(49.4Kd),correspondingtotheN-proteinmolecularweight.Thededucedproteinpossesses43serineresidues(9.6%ofthetotalaminoacidcontent)whichmaybephosporylatedandinvolvedinN-protein/RNAbinding.N-proteinalsohas5r
7、egionswithahighbasicaminoacidcontent.Oneofthemisalsoserine-richandhasastronghomologysitewithMHV,TGEandIBVviruses.InthefirstpartoftheN-terminal,a12-amino-acidsequence(PRWYFYYLGTGP)ishighlyconservedforBECV-F15,JHM,TGEandIBVviruses.BCVMebusstrai
