携带卵清蛋白t细胞表位的兔出血症病毒vp60蛋白病毒样颗粒的表达与其免疫原性分析

《携带卵清蛋白t细胞表位的兔出血症病毒vp60蛋白病毒样颗粒的表达与其免疫原性分析》由会员上传分享，免费在线阅读，更多相关内容在学术论文-天天文库。

1、ABSTRACTCoNSTRUCTIoNANDIMMUNoGENICITYoFCHIMERICPROTEINSCONSTRUCTEDBYOVATCELLEPITOPEINSERTEDINTOTHECAPSIDPROTEINVP60OFRABBITHEMORRHAGICDISEASEⅥRUSRabbitHemorrhagicDisease(RHD)，causedbyRabbithaemorrhagicdiseasevirus，ischaracterizedbyacute，highlycontagious，systemicsolidorganbleedingandalarge

2、areaofdeathinrabbits．RHDV,amemberofthecalicivirus,couldinducestronghumeral，cellularandmucosalinlmuneresponsesinrabbits．VP60isamajorviralstructuralprotein，whichcouldberelatedtoanti—viraliffmluneresponseduringRHDVinfection．ThecapsidproteinofRHDVspontaneouslyassemblesintoVLPwhicharemorpholog

3、icallyandantigenicallyindistinguishablefromnativevirions，butdevoidoftheviralRNA．PreviousresearcheshavealreadydemonstratedthatnonadjuvantedVLPscouldinducerobusthumeralandcell-mediatedimmuneresponses．MorerecentresearchhasalsoindicatedthatRHDVmightbeusedasanimmunestimulatorandself-adjuvant．R

4、esearcheshaveshownthatVLPscouldbeexcellentcandidatestoinduceapotentimmuneresponsetoforei印antigensinsertedintheirparticulatestructureandtobeavaccinevector．Inordertoexplorethecapacityofrabbithemorrhagicdiseasevirus—likeparticles(RHDV-VLPs)accommodatingexogenousantigenepitopes，theT-cellepito

5、pe(fragmentofovalbumin，OVA)WasembeddedintoRHDVVLPs．ThechimericproteinsgeneratedweretestedforassemblyintoVLPs，forantigenicityandimmunogenicity,includingtheircapacitytoinduceimmuneresponses，andfortheircapacitytopresenttheforeignepitope．Majortestsandresultsaredescirbedasfollow：1．Construction

6、andidentifieationoftherecombinantshuttlevectorsTheCD8+Tcellepitope(SIINFEKL)wereinsertedintothedifferentsitesofRHDVVLPs：1)directlyinsertedintotheVP60C-terminal；2)replacedtheVP60502—510aminoacidofVP60；3)replacedtheVP60411-417aminoacidofVP60；4)replacedtheVP60302-309aminoacidofVP60．Thesequen

7、cescodingchimericproteinswereobtainedbyPCRorSOEPCR．ThePCRproductwasdigestedandclonedintocorrespondingenzymedigestedpFastBacTMHT1plasmid．ThefourrecombinanttransfervectorsweretransformedIII携带卵清蛋白(OVA)片段的兔出血症病毒VP60蛋自病毒样颗粒的表达及其免疫原性研究intoE．coliDHlOBac．nlecoloniesofE．coli