Molecular Basis of Rare Aminoglycoside Susceptibility and Pathogenesis of Burkholderia pseudomallei Clinical Isolates from Thailand 英文参考文献.docVIP

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Molecular Basis of Rare Aminoglycoside Susceptibility and Pathogenesis of Burkholderia pseudomallei Clinical Isolates from Thailand 英文参考文献.doc

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Molecular Basis of Rare Aminoglycoside Susceptibility and Pathogenesis of Burkholderia pseudomallei Clinical Isolates from Thailand 英文参考文献

MolecularBasisofRareAminoglycosideSusceptibility andPathogenesisofBurkholderiapseudomalleiClinical IsolatesfromThailand LilyA.Trunck1,KatieL.Propst1,VanapornWuthiekanun2,ApichaiTuanyok3,StephenM.Beckstrom- Sternberg3,4,JamesS.Beckstrom-Sternberg3,SharonJ.Peacock2,5,PaulKeim3,4,StevenW.Dow1, HerbertP.Schweizer1* 1DepartmentofMicrobiology,ImmunologyandPathology,RockyMountainRegionalCenterofExcellenceforBiodefenseandEmergingInfectiousDiseasesResearch, ColoradoStateUniversity,FortCollins,Colorado,UnitedStatesofAmerica,2FacultyofTropicalMedicine,MahidolUniversity,Bangkok,Thailand,3TheMicrobialGenetics andGenomicsCenter,NorthernArizonaUniversity,Flagstaff,Arizona,UnitedStatesofAmerica,4PathogenGenomicsDivision,TranslationalGenomicsResearchInstitute, Flagstaff,Arizona,UnitedStatesofAmerica,5DepartmentofMedicine,UniversityofCambridge,Addenbrooke’sHospital,Cambridge,UnitedKingdom Abstract Background:Burkholderiapseudomalleiisintrinsicallyresistanttoaminoglycosidesandmacrolides,mostlyduetoAmrAB- OprA efflux pump expression. We investigated the molecular mechanisms of aminoglycoside susceptibility exhibited by Thaistrains708a,2188a,and3799a. Methodology/PrincipalFindings:qRT-PCRrevealedabsenceofamrBtranscriptsin708aandgreatlyreducedlevelsin2188a and 3799a. Serial passage on increasing gentamicin concentrations yielded 2188a and 3799a mutants that became simultaneouslyresistanttootheraminoglycosidesandmacrolides,whereassuchmutantscouldnotbeobtainedwith708a. Transcript analysis showed that the resistance of the 2188a and 3799a mutants was due to upregulation of amrAB-oprA expressionbyunknownmechanism(s).UseofaPCRwalkingstrategyrevealedthattheamrAB-oprAoperonwasmissingin 708aandthatthislosswasassociatedwithdeletionofmorethan70kbofgeneticmaterial.RescueoftheamrAB-oprBregion from a 708a fosmid library and sequencingshowed thepresence of a largechromosome1 deletion(131kb and 141kb comparedtostrainsK96243and1710b,respectively).ThisdeletionnotonlyremovedtheamrAB-oprAoperon,butalsothe