Functional Characterization of a Lipoprotein-Encoding Operon in Campylobacter jejuni 英文参考文献.docVIP

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Functional Characterization of a Lipoprotein-Encoding Operon in Campylobacter jejuni 英文参考文献.doc

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Functional Characterization of a Lipoprotein-Encoding Operon in Campylobacter jejuni 英文参考文献

FunctionalCharacterizationofaLipoprotein-Encoding OperoninCampylobacterjejuni MayumiOakland¤a,ByeonghwaJeon¤b,OrhanSahin,ZhangqiShen,QijingZhang* DepartmentofVeterinaryMicrobiologyandPreventiveMedicine,IowaStateUniversity,Ames,Iowa,UnitedStatesofAmerica Abstract Background: Bacteriallipoproteinshaveimportantfunctionsinbacterialpathogenesisandphysiology.In Campylobacter jejuni, a major foodborne pathogen causing gastroenteritis in humans, the majority of lipoproteins have not been functionallycharacterized.Previously,weshowedbyDNAmicroarraythatCmeR,atranscriptionalregulatorrepressingthe expressionofthemultidrugeffluxpumpCmeABC,modulatestheexpressionofathree-geneoperon(cj0089,cj0090 ,and cj0091)encodingaclusteroflipoproteinsinC.jejuni. Methodology/Principal Findings: In this work, we characterized the function and regulation of the cj0089-cj0090-cj0091 operon.IncontrasttotherepressionofcmeABC,CmeRactivatestheexpressionofthelipoproteingenesandtheregulation isconfirmedbyimmunoblottingusinganti-Cj0089andanti-Cj0091antibodies.GelmobilityshiftassayshowedthatCmeR directlybindstothepromoterofthelipoproteinoperon,butthebindingismuchweakercomparedwiththepromoterof cmeABC.AnalysisofdifferentcellularfractionsindicatedthatCj0089wasassociatedwiththeinnermembrane,whileCj0091 waslocatedontheoutermembrane.Inactivationofcj0091,butnotcj0089,significantlyreducedtheadherenceofC.jejuni to INT 407 cells in vitro, indicating that Cj0091 has a function in adherence. When inoculated into chickens, the Cj0091 mutant also showed a defect in early colonization of the intestinal tract, suggesting that Cj0091 contributes to Campylobactercolonizationinvivo.ItwasalsoshownthatCj0091wasproducedandimmunogenicinchickensthatwere naturallyinfectedbyC.jejuni. Conclusion/Significance:TheseresultsindicatethatthelipoproteinoperonissubjecttodirectregulationbyCmeRandthat Cj0091functionsasanadhesionmechanisminC.jejuniandcontributestoCampylobactercolonizationoftheintestinaltract inanimalhosts. Citation: O