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1、J.gen.Virol.(1985),66,719726.PrintedinGreatBritain719Keywords:coronavirus1BV/spikeprotein/DNAsequence/mRNAECloningandSequencingoftheGeneEncodingtheSpikeProteinoftheCoro~:virusIBVByMATTHEWM.BINNS,*MICHAELE.G.BOURSNELL,DAVIDCAVANAGH,DARRYLJ.C.PAPPIN1ANDT.DAVIDK.BROWNH
2、oughtonPoultryResearchStation,Houghton,Huntingdon,Cambs.PE172DAand1DepartmentofBiochemistry,Unh'ersityofLeeds,LeedsLS29JT,U.K.(Accepted19December1984)SUMMARYRNAsequencesencodingthesurfaceprojection(spike)ofthecoronavirusinfectiousbronchitisvirus,strainBeaudette,have
3、beenclonedintopBR322usingcDNAprimedwithaspecificoligonucleotide.A5.3kilobaseviralinsertintheclonepMB179hasbeenidentified.Theregionofthisclonecodingforthespikegenehasbeensequencedbythechainterminationmethod,andwepresentherethefirstreportofDNAsequencedataforacoronavir
4、usspikeprotein,theproteinwhichformsthecharacteristic'corona'afterwhichthegroupisnamed.Theaminoacidsequenceoftheprimarytranslationproduct,deducedfromtheDNAsequence,predictsapolypeptideof1162aminoacidswithamolecularweightof127006.Thishasmanyinterestingfeatureswhichcon
5、firmandextendourknowledgeofthisrecentlycharacterizedmembraneglycoprotein.ThepolypeptideissubsequentlycleavedtoS1and$2,andpartialaminoacidanalysisoftheamino-terminusoftheS1polypeptidehasbeenemployedtolocatethepositionofthisterminusofS1withinthelargeopenreadingframe.T
6、heaminoacidanalysisalsorevealsthepresenceofan18aminoacidputativesignalsequenceontheprimarytranslationproductwhichisnotpresentonthematureS1polypeptide.INTRODUCTIONInfectiousbronchitisvirus(IBV)causesrespiratorydiseaseinthefowlandisofconsiderableeconomicimportancetoth
7、epoultryindustry.ThetypespeciesoftheCoronaviridae,itpossessesasingle-strandedRNAgenome,approximately20kbinlength,ofpositivepolaritywhichspecifiestheproductionofthreemajorstructuralproteins:nucleocapsidprotein,membraneglycoprotein,andspikeglycoprotein.Thespikeprotein
8、,encodedbymRNAE,hasrecentlybeencharacterized(Cavanagh,1983a,b,c)ascomprisingtwoorthreecopieseachoftwoglycopolypeptides,$1(90000mol.wt.)and
