马传染性贫血病毒弱毒疫苗致弱过程病毒基因的进化分析-evolutionary analysis of the genes of equine infectious anemia virus attenuated vaccine-induced attenuated process virus.docx

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马传染性贫血病毒弱毒疫苗致弱过程病毒基因的进化分析-evolutionary analysis of the genes of equine infectious anemia virus attenuated vaccine-induced attenuated process virus.docx

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马传染性贫血病毒弱毒疫苗致弱过程病毒基因的进化分析-evolutionary analysis of the genes of equine infectious anemia virus attenuated vaccine-induced attenuated process virus

236D/-,237N/K,247E/Kand321K/Eingp90,7R/HinTat,41T/I,51T/Iand55Q/KinS2,74V/IinRevandchangeoftranscription-factorAP-1bindingmotifinthenegativeregulatingelement(NRE),transcriptionfactorbindingmotifE-boxwithintheenhancer,thetranscriptionstartpositionandthefirstresidueoftrans-activationresponsive(TAR)elementofLTR.(2)PhylogeneticanalysisshowedthatthesequencesofvirusesisolatedfromEIAVLN40-infectedhorseswereobviouslysplitintotwomajorbranches.VirusesisolatedintheearlystageandintheasymptomaticstageofinfectioninallEIAVLN40-infectedhorsesclusteredintoabranchedifferentfromEIAVLN40.VirusesisolatedafterdiseaseonsetremainedinthesamebranchasEIAVLN40.Thevariationsofgp90weremostlyconcentratedineighthighlyvariableregionsV1-V8.ChangesintheglycosylationsiteswithinV3,V4andV5regionswereusuallyassociatedwiththediseasestatus.Specifically,glycosylationsites191NSSN194,237NNTW240,and280NDTS283withinthesethreeregionswerepresentinEIAVLN40andmostofthequasispeciesisolatedafter,butnotbefore,diseaseonset.DuringtheevolutionofEIAVLN40,over12%substitutionsofaminoacidresiduespresentedinS2,whichassociatedwiththediseasestatus.(3)Phylogeneticanalysisshowedthatthegp90sequencesofvirusesisolatedfromEIAVDLV121-infectedhorseswereobviouslysplitintothreemajorbranches.Thedivergencesofgp90sequencesisolatedfromEIAVDLV121-inoculatedhorseswere3.88%-6.73%comparedwithEIAVLN40,andwere0.91%-6.49%whencomparedwithEIAVDLV121.TheS2sequencesofvirusesisolatedfromEIAVDLV121-infectedhorseswereobviouslysplitintotwobranches,theaveragedifferencesbetweentheS2sequencesfromEIAVDLV121-inoculatedhorsesandEIAVLN40werefrom0.79%-8.83%,andwere3.08%-8.14%whencomparedwithEIAVDLV121.Conclusion:AseriesofconsensussubstitutionsinmultiplegenesoftheattenuatedEIAVvaccinestrainweregeneratedduringattenuationprocess.Themutationsingp90andS2wereassociatedwiththediseasestatus.Thevaccinestrainreplicatesinarelativelylowlevelininoculatedhorses,whichenabledtothevaccinequasispeciestoevolvecontinuousandtoincreasediversityofgp90.KeyWords:Equineinfectiousanemia;Equ