Mycolactone Gene Expression Is Controlled by Strong SigA-Like Promoters with Utility in Studies of Mycobacterium ulcerans and Buruli Ulcer 英文参考文献.docVIP

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Mycolactone Gene Expression Is Controlled by Strong SigA-Like Promoters with Utility in Studies of Mycobacterium ulcerans and Buruli Ulcer 英文参考文献.doc

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Mycolactone Gene Expression Is Controlled by Strong SigA-Like Promoters with Utility in Studies of Mycobacterium ulcerans and Buruli Ulcer 英文参考文献

MycolactoneGeneExpressionIsControlledbyStrong SigA-LikePromoterswithUtilityinStudiesof MycobacteriumulceransandBuruliUlcer NicholasJ.Tobias1,2,TorstenSeemann3,SachaJ.Pidot1,2,JessicaL.Porter2,LaurentMarsollier4 ,Estelle Marion4,5,FranckLetournel6,TasnimZakir1,JosephAzuolas7,JohnR.Wallace8,HuiHong9,JohnK. Davies1,BenjaminP.Howden1,2,10,PaulD.R.Johnson10,GrantA.Jenkin1,2,TimothyP.Stinear1,2 * 1Department of Microbiology, Monash University, Clayton, Victoria, Australia, 2Victorian Bioinformatics Consortium, Monash University, Clayton, Victoria, Australia, 3DepartmentofMicrobiologyandImmunology,UniversityofMelbourne,Parkville,Victoria,Australia,4Grouped’EtudedesInteractionsHo?te-Pathoge`ne,UPRES-EA3142, Universite′ d’Angers, Angers, France, 5Laboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire, Angers, France, 6Laboratoire de Neurobiologie et Transge′ne`se,UPRES-EA 3143,Universite′ d’Angers, Angers, France, 7Department ofPrimaryIndustries,Mickleham Road,Attwood,Victoria, Australia, 8Departmentof Biology,MillersvilleUniversity,Millersville,Pennsylvania,UnitedStatesofAmerica,9DepartmentofBiochemistry,UniversityofCambridge,Cambridge,UnitedKingdom, 10DepartmentofInfectiousDiseases,AustinHealth,Heidelberg,Victoria,Australia Abstract Mycolactone A/B is a lipophilic macrocyclic polyketide that is the primary virulence factor produced by Mycobacterium ulcerans,ahumanpathogenandthecausativeagentofBuruliulcer.InM.ulceransstrainAgy99themycolactonepolyketide synthase(PKS)locusspansa120kbregionofa174kbmegaplasmid.HerewehaveidentifiedpromoterregionsofthisPKS locususingGFPreporterassays,insilicoanalysis,primerextension,andsite-directedmutagenesis.Transcriptionofthelarge PKS genes mlsA1 (51kb), mlsA2 (7kb) and mlsB (42kb) is driven by a novel and powerful SigA-like promoter sequence situated 533bp upstream of both the mlsA1 and mlsB initiation codons, which is also functional in Escherichia coli, Mycobacterium smegmatis and Mycobacterium marinum. Promoter regions