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1、VirusResearch,9(1988)343-356343ElsevierVRR00394TemporalregulationofbovinecoronavirusRNAsynthesisJamesG.Keck,BrendaG.Hogue*,DavidA.Brian**andMichaelM.C.LaiDepartmentsofMicrobiologyandNeurology,UniversityofSouthernCalifornia,SchoolofMedicine,LasAngeles,California,U.S.A.andDepartmentofMicrobi
2、ology,UniversityofTennessee,Knoxville,Tennessee,U.S.A.(Acceptedforpublication9November1987)Thestructureandsynthesisofbovinecoronavirus(BCQspecificintracellularRNAwerestudied.Agenome-sizeRNAandsevensubgenomicRNAswithmolecu-larweightsofapproximately3.3X106,3.1X106,2.6X106,1.1X106,1.0X106,0.8
3、Xlo6and0.6xlo6weredetected.ComparisonsofBCVintracellularRNAswiththoseofmousehepatitisvirus(MHV)demonstratedthepresenceofanadditionalRNAforBCV,species2a,of3.1xlo6daltons.BCVRNAscontainanested-setstructuresimilartothatofothercoronaviruses.Thisnested-setstructurewouldsuggestthatthenewRNAhasac
4、apacitytoencodeaproteinofapproxi-mately430aminoacids.Kineticstudiesdemonstratedthatthesynthesisofsubge-nomicmRNAsandgenomicRNAaredifferentiallyregulated.At4to8hpost-infection(p.i.),subgenomicRNAsaresynthesizedatamaximalrateandrepresentgreaterthan90%ofthetotalviralRNAsynthesized,whereasgeno
5、me-sizeRNAaccountsforonly7%.Laterininfection,at70to72hp.i.,genome-sizeRNAismuchmoreabundantandaccountsfor88%oftotalRNAsynthesized.Im-munoprecipitationsof[35S]methionine-pulse-labeledviralproteinsdemonstratedthatviralproteinsynthesisoccursearlyintheinfection,concurrentwiththepeakofviralsubg
6、enomicRNAsynthesis.Westernblotanalysissuggeststhattheseproteinsarestablesincetheproteinsarepresentathighlevelaslateas70to72hp.i.TheCorrespondenceto:J.G.Keck,Depts.ofMicrobiologyandNeurology,UniversityofSouthernCalifornia,SchoolofMedicine,LosAngeles,CA90033-1054,U.S.A.*Currentaddress:Depart
7、mentofMicrobiologyandImmunology,UniversityofSouthernCaliforniaatLosAngeles,SchoolofMedicine,LosAngeles,CA90024,U.S.A.344kineticsofproductionofvirusparticlescoincideswiththesynthesisofgenomicRNA.Thesestudiesthusindicatethatthereisadifferentialtemporalregulation