Analysis of an Ordered, Comprehensive STM Mutant Library in Infectious Borrelia burgdorferi Insights into the Genes Required for Mouse Infectivity 英文参考文献.docVIP
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Analysis of an Ordered, Comprehensive STM Mutant Library in Infectious Borrelia burgdorferi Insights into the Genes Required for Mouse Infectivity 英文参考文献
AnalysisofanOrdered,ComprehensiveSTMMutant
LibraryinInfectiousBorreliaburgdorferi:Insightsinto
theGenesRequiredforMouseInfectivity
TaoLin1*,LihuiGao1,ChuhuaZhang1,EvelynOdeh1,MaryB.Jacobs3,Lo?¨cCoutte1¤,GeorgeChaconas4,
MarioT.Philipp3,StevenJ.Norris1,2
1DepartmentofPathologyandLaboratoryMedicine,MedicalSchool,UniversityofTexasHealthScienceCenteratHouston,Houston,Texas,UnitedStatesofAmerica,
2DepartmentofMicrobiologyandMolecularGenetics,MedicalSchool,UniversityofTexasHealthScienceCenteratHouston,Houston,Texas,UnitedStatesofAmerica,
3Department of Parasitology, Tulane Regional Primate Research Center, Tulane University Health Sciences Center, Covington, Louisiana, United States of America,
4DepartmentofBiochemistryandMolecularBiologyandDepartmentofMicrobiologyandInfectiousDiseases,TheUniversityofCalgary,Calgary,Alberta,Canada
Abstract
TheidentificationofgenesimportantinthepathogenesisofLymediseaseBorreliahasbeenhamperedbyexceedinglylow
transformation rates in low-passage, infectious organisms. Using the infectious, moderately transformable B. burgdorferi
derivative5A18NP1andsignature-taggedversionsoftheHimar1transposonvectorpGKT,wehaveconstructedadefined
transposon library for the efficient genome-wide investigation of genes required for wild-type pathogenesis, in vitro
growth, physiology, morphology, and plasmid replication. To facilitate analysis, the insertion sites of 4,479 transposon
mutantsweredeterminedbysequencing.ThetransposoninsertionswerewidelydistributedacrosstheentireB.burgdorferi
genome, with anaverage of 2.68unique insertion sites per kb DNA. The 10 linear plasmids and 9 circular plasmids had
insertionsin33to100percentoftheirpredictedgenes.Incontrast,only35%ofgenesinthe910kblinearchromosome
hadincapacitatinginsertions;therefore,theremaining601chromosomalgenesmayrepresentessentialgenecandidates.In
initialsignature-taggedmutagenesis(STM)analyses,434mutantswereexaminedatmultipletissuesitesforinfectivityin
miceusingasemi-quantitative,Luminex-basedDNAdetect
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