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1、Volume94,number2FEBSLETTERSOctober1978THEGENOMESOFTHREECORONAVIRUSESMalcolmR.MACNAUGHTONDivisionofCommunicableDiseases,ClinicalResearchCentre,Harrow,MiddlesexHAISlJJ,EnglandReceived11August19781.Introductionresultsshowsamuchgreatersimilaritybetweencoronavirusgenom
2、esthanhasbeenpreviouslyAnumberofstudiesonthegenomicRNAsofobserved.coronaviruseshavebeenreported.Thegenomeofavianinfectiousbronchitisvirus(IBV)isasinglesingle-strandedmoleculeofhighmol.wt[l-4],2.Materialsandmethodswhichispronetodegradationintosmallermolecules[1,5].
3、Thereissomedisagreementconcerningitsmol.wtwithestimatesvaryingfrom9.0Xlo6[l]IBVBeauwasgrowninconfluentprimarychickto5.6Xlo6[4].Similarstudiesonthehumankidneycellculturesandlabelledwith[3H]uridineorcoronavirus(HCV)genomehavealsoshownittobeat3H]adenosineasin[3].HCV2
4、29Ewasgrowninlargemoleculeofmol.wtvaryingfrom6.1Xlo6forconfluentmonolayerculturesofcontinuousMRCstrainOC43[6]to5.8XIO6forstrain2298[7].cellsandlabelledwith[3H]uridineor[3H]adenosineAgain,aswithIBV,theHCVgenomeisproneto[7].Finally,MHV3wasgrowninconfluentsecondaryde
5、gradation[6,7].Thegenomesoftwoporcinemouseembryonicfibroblasts.Thecellmonolayerscoronaviruses,transmissiblegastroenteritisviruswereinfectedataninputmultiplicityof0.1infectious(TGEV)andhaemagglutinatingencephalomyelitisparticlespercell,withanadsorptionperiodof1.5hv
6、irus(HEV),havealsobeenshowntobelargesingleat37C.At6hafterinfection,20pCi/mloft3H]-single-strandedRNAmoleculesofabout60Sthaturidineor[3H]adenosine(RadiochemicalCentre,dissociateinto35Sand4SmaterialonheatingaboveAmersham)wereaddedtothemedium.At32hafter60°C[8]~Corona
7、virusgenomespossesscertaininfectionthecellswereharvestedandsubjectedtomRNAcharacteristics.Polyadenylicacid[poly(A)]3freeze-thawcycles.TheviruspreparationsweresequencesarefoundinthegenomesofIBV[2,4,9]purifiedonsucrosegradientsandlabelledvirusandHCV[7,10]atornearthe
8、ir3-termini[7].particleswithtypicalcoronavirusmorphologywereFurthermore,thegenomeofIBVhasbeenshowntoobtainedinpeakfractionsofdensity1.I8g/mlbeinfectious