Differential Gene Expression and Adherence of Escherichia coli O157H7 In Vitro and in Ligated Pig Intestines 英文参考文献.docVIP

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Differential Gene Expression and Adherence of Escherichia coli O157H7 In Vitro and in Ligated Pig Intestines 英文参考文献.doc

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Differential Gene Expression and Adherence of Escherichia coli O157H7 In Vitro and in Ligated Pig Intestines 英文参考文献

DifferentialGeneExpressionandAdherenceof EscherichiacoliO157:H7InVitroandinLigatedPig Intestines XianhuaYin1,4,JingZhu2,YanniFeng3,JamesR.Chambers1,JoshuaGong1,CarltonL.Gyles4* 1GuelphFoodResearchCenter,AgricultureandAgri-FoodCanada,Guelph,Ontario,Canada,2CollegeofLifeScienceandBiotechnology,ShanghaiJiaoTongUniversity, Shanghai,People’sRepublicofChina,3CollegeofAnimalScienceandTechnology,QingdaoAgriculturalUniversity,Qingdao,People’sRepublicofChina,4Departmentof Pathobiology,OntarioVeterinaryCollege,UniversityofGuelph,Guelph,Ontario,Canada Abstract Background:EscherichiacoliO157:H7strain86–24growninMacConkeybroth(MB)showsalmostnoadherencetocultured epithelial cells but adheres well in pig ligated intestines. This study investigated the mechanisms associated with the differencebetweenin-vitroandin-vivoadherenceoftheMBculture. Methodology/PrincipalFindings:ItwasfoundthatdecreasedadherenceinvitrobybacteriagrowninMBwasmainlydue to lactose, possibly implicating the involvement of carbon catabolite repression (CCR). Expression of selected virulence- relatedgenesassociatedwithadherenceandCCRwasthenexaminedbyquantitativePCR.Whenbacteriaweregrownin MB and Brain Heart Infusion with NaHCO3 (BHIN) plus lactose, pH was reduced to 5.5–5.9 and there was a significant decreaseinexpressionofthelocusofenterocyteeffacement(LEE)geneseae,tir,espD,grlA/Randler,andanincreaseincya (cAMP),andtwonegativeregulatorsoftheLEE,gadEandhfq.PutativevirulencegenesstcE,hlyA,entandnleAwerealso decreasedinvitro.Reversalofthesechangeswasnotedforbacteriarecoveredfromtheintestine,wheretranscriptsforqseF andfisandputativevirulencefactorsAidA15,TerCandEnt/EspL2weresignificantlyincreased,andtranscriptsforAIDA48 ,Iha, UreC,Efa1A,Efa1B,ToxB,EhxA,StcE,NleAandNleBwereexpressedathighlevels. Conclusions/Significance: Presence of lactose resulted in decreased expression of LEE genes and the failure of EHEC O157:H7 to adhere to epithelial cells in vitro but this repression was overcome in vivo. CCR and/or acidic pH