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1、VeterinaryMicrobiology,21(1989)115-123115ElsevierSciencePublishersB.V.,Amsterdam--PrintedinTheNetherlandsPurificationofInfectiousBronchitisCoronavirusbySephacrylS-1000GelChromatographyH.NAGANO1,K.YAGYU2andS.OHTA21ResearchCenterforVeterinaryScienceofTheKitasatoInstitute,Kashiw
2、a,Chiba(Japan)2NationalVeterinaryAssayLaboratory,Kokubunji,Tokyo(Japan)(Acceptedforpublication5June1989)ABSTRACTNagano,H.,Yagyu,K.andOhta,S.,1989.PurificationofinfectiousbronchitiscoronavirusbySephacrylS-1000gelchromatography.Vet.Microbiol.,21:115-123.Aprocedurewasdevelopedto
3、purifyinfectiousbronchitisvirus(IBV)bygelchromatography(GC)withaSephacrylS-1000column.VirussamplesconcentratedbycentrifugationwereappliedtoaSephacrylS-1000columnandelutedby0.02Mphosphatebuffer(pH7.2)containing0.15MNaC1.Virusparticleswererecoveredmainlyinthefirstpeak.Purityoft
4、hesampleswasevaluatedbybothsodiumdodecylsulfate-polyacrylamidegelelectrophoresisandelectronmicroscopy.Usingelectronmicroscopy,itwasfoundthatthereweremorespike-richparticlesinthevirussamplespurifiedbyGCthaninthosepurifiedbysucrosedensitygradientcentrifugation(SDGC).Inaddition,
5、thehemagglutinationunit[logzo(infectivitytiter/hemagglutinationtiter)]ofGC-purifiedvirussampleswas~10timeslowerthanthatofSDGC-purifiedvirussamples.TheseresultsindicatethatSephacrylS-1000gelchromatographyisusefulforpurificationofIBV.INTRODUCTIONInfectiousbronchitisvirus(IBV)be
6、longstotheCoronaviridae.TheIBVvirionwithadiameterof~120nmisenvelopedandcontainsonemoleculeofaninfectious(plus-sense),single-strandedRNA(RobbandBond,1979;Shid-delletal.,1982).Thevirionismainlycomposedofspike(S;peplomer)withamolecularweightof128kDa,membrane(M)withamolecularweig
7、htof45kDaandnucleocapsid(N)withamolecularweightof25kDa(Spaanetal.,1988).SandMproteinsarebothglycosylatedandpartsofthemareexposedatthesurfaceofthevirion.Sproteinconsistsoftwoglycoproteins,S1and$2(Cavanagh,1981,1983;Sternetal.,1982).TheSproteinofIBVmayplayanimportantroleinviral
8、activities,suchashemagglutination(HA),infectivityforculturedcells,an
