Generation of Leishmania Hybrids by Whole Genomic DNA Transformation 英文参考文献.docVIP

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Generation of Leishmania Hybrids by Whole Genomic DNA Transformation 英文参考文献.doc

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Generation of Leishmania Hybrids by Whole Genomic DNA Transformation 英文参考文献

GenerationofLeishmaniaHybridsbyWholeGenomic DNATransformation AdrianoC.Coelho,PhilippeLeprohon,MarcOuellette* CentredeRechercheenInfectiologie,Universite′ Laval,Que′bec,Canada Abstract Geneticexchangeisapowerfultooltostudygenefunctioninmicroorganisms.Here,wetestedthefeasibilityofgenerating LeishmaniahybridsbyelectroporatinggenomicDNAofdonorcellsintorecipientLeishmaniaparasites.ThedonorDNAwas markedwithadrugresistancemarkerfacilitatingtheselectionofDNAtransferintotherecipientcells.ThetransferredDNA was integrated exclusively at homologous locus and was as large as 45kb. The independent generation of L. infantum hybrids with L. major sequences was possible for several chromosomal regions. Interfering with the mismatch repair machinerybyinactivatingtheMSH2geneenabledanincreasedefficiencyofrecombinationbetweendivergentsequences, hencefavouringtheselectionofhybridsbetweenspecies.Hybridswereshowntoacquirethephenotypederivedfromthe donorcells,asdemonstratedforthetransferofdrugresistancegenesfromL.majorintoL.infantum.Thedescribedmethod is a first step allowing the generation of in vitro hybrids for testing gene functions in a natural genomic context in the parasiteLeishmania. Citation:CoelhoAC,LeprohonP,OuelletteM(2012)GenerationofLeishmaniaHybridsbyWholeGenomicDNATransformation.PLoSNeglTropDis6(9):e1817. doi:10.1371/journal.pntd.0001817 Editor:ElodieGhedin,UniversityofPittsburgh,UnitedStatesofAmerica ReceivedJune19,2012;AcceptedAugust1,2012;PublishedSeptember20,2012 Copyright: ? 2012 Coelho et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:ThisworkwassupportedbyCanadianInstitutesofHealthResearch(CIHR)operatinggrants(toM.O.)andapostdoctoralfellowshipfromtheCIHR-STP StrategicTrainingprogram(toA.C.C.).FundingfortheopenaccesschargewasprovidedbyCanadianInstitutesofHealthResearchGrant(M.O.).T