identification of the structural proteins of vp1 and vp2

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1、JournalofVirologicalMethods171(2011)323–328ContentslistsavailableatScienceDirectJournalofVirologicalMethodsjournalhomepage:www.elsevier.com/locate/jvirometProtocolsIdentificationofthestructuralproteinsofVP1andVP2ofanovelmudcrabdicistrovirusRuiZhang1,JianguoHe1,Hongju

2、nSu,ChuanfuDong,ZhixunGuo,YujieOu,XiexiongDeng,ShaopingWeng∗StateKeyLaboratoryforBiocontrol,SchoolofLifeSciences,SunYat-Sen(Zhongshan)University,135XingangRoadWest,Guangzhou510275,ChinaabstractArticlehistory:Mudcrabdicistrovirus(MCDV),anewlyidentifiedsingle-strandedp

3、ositiveRNAvirus,isanimportantReceived25February2010pathogenthatcausesseriouseconomiclossestomudcrabaquaculture.Inthisstudy,MCDVwaspuri-Receivedinrevisedform24August2010fied,andthreestructuralproteinsofMCDVwereseparatedbySDS-PAGE.TheN-terminal15aminoacidsAccepted8Sept

4、ember2010weresequencedandalignedwiththemainstructuralproteinsofotherdicistrovirus.ThethreestructuralAvailableonline17September2010proteinswerenamedVP1,VP2andVP3.Monoclonalantibodies(MAbs)againstthetwomainstructuralproteins,VP1andVP2,wereprepared,andthetwostructuralp

5、roteinswerethenidentifiedusingtheseKeywords:MAbs.TheresultsofWesternblotanalysesdemonstratedthatfiveMAbsrecognisedVP1andtworecog-Mudcrabdicistrovirus(MCDV)nisedVP2.Theresultsofimmunogoldtransmissionelectronmicroscopy(IEM)revealedthattheepitopesStructuralproteinMonoclo

6、nalantibodies(MAbs)ofthetwostructuralproteinsrecognisedbytheMAbswerelocatedattheoutersurfaceofthevirions,whichsuggestedthatthetwostructuralproteinsareMCDVcapsidproteins.TheidentificationofthetwostructuralproteinsofMCDVisusefulforstudyingtheirfunctions,aswellasthemech

7、anismofinfectionandthepathogenesisofMCDV.©2010ElsevierB.V.Allrightsreserved.1.IntroductionsequenceswithTSVandotherdicistroviruses,thecharacteristicsofthestructuralproteinsindicatethatitbelongstotheviralfam-Themudcrab(Scyllaserrata),animportantspecieseconom-ilyDicist

8、roviridae.Thisviruswasnamedmudcrabdicistrovirusically,iscultivatedcommerciallyinmanycountries,including(MCDV).China,IndiaandAustralia(Cele