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1、J.gen.Virol.(I978),39,497-504497PrintedinGreatBritainTheGenomeofHumanCoronavirusStrain229EByM.R.MACNAUGHTONANDM.HILARYMADGEDivisionofCommunicableDiseases,ClinicalResearchCentre,Harrow,MiddlesexHAI3UJ,U.K.(Accepted23December1977)SUMMARYThegenomicRNAofhumancoronavirusstrain229E(HCV229E)migratedonp
2、olyacrylamidegelsasasinglepeakwithatool.wt.of5"8xIO6.Denaturationofthegenomewithformaldehydedidnotalteritselectrophoreticmobility,whichsuggeststhattheI-ICVEe9Egenomeisasingle-strandedmolecule.Atleast3o~ofthegenomicRNAwasshowntocontaincovalentlyattachedpolyadenylicacid[poly(A)]sequencesbybindingt
3、heRNAtoanoligo(dT)-cellulosecolumn.Thesepoly(A)tractswereshowntobeabout7onucleotidesinlengthbymeasuringtheresistancetodigestionofHCV229ERNAwithpancreaticandT1RNases.Finally,thegenomicRNAwasshowntoterminateatornearthe3'-terminusonthebasisofitssusceptibilitytopolynucleotidephosphorylase.INTRODUCTI
4、ONHumancoronavirusstrain229E(HCV229E)isclassifiedasamemberofthecorona-viruses(Tyrrelletal.t968).Thecoronavirusesaregroupedtogetherprimarilyontheirsimilarmorphologyinthattheyarealllipid-containingenvelopedvirusesrangingindiam.from8otoI2onmandpossesswidely-spacedclub-shapedsurfaceprojectionsupto2o
5、nminlength(Mclntosh,I974).Thepropertiesofthegenomesofcertaincoronaviruseshavebeenreported.Avianinfectiousbronchitisvirus(IBV)(Watkinsetal.I975;Lomniczi,I977;Macnaughton&Madge,I977,Schochetmanetal.I977),transmissiblegastroenteritisvirus(TGEV)(Garwesetal.I975)andHCVstrainOC43(Tannock&Hierholzer,i9
6、77)havebeenshowntoconsistofalargemol.wt.singlesingle-strandedRNAmolecule.Furthermore,thegenomeofIBVhasbeenshowntocontaincovalentlyattachedpoly-adenylicacid[poly(A)]sequences(Lomniczi,1977;Macnaughton&Madge,1977;Schochet-manetal.I977).InthispaperwehaveshownthattheHCV229Egenomealsoexistsasasingles
7、ingle-strandedRNAmoleculeoftool.wt.about5"8xio6withatleast3o%ofthemoleculescontainingpoly(A)sequencesofabout7onucleotidesinlength,locatedatornearthe3'-terminusofthemolecule.METHODSVirusculture.HCVstrain229EwasobtainedfromDrS
