FRUIT, a Scar-Free System for Targeted Chromosomal Mutagenesis, Epitope Tagging, and Promoter Replacement in Escherichia coli and Salmonella enterica 英文参考文献.docVIP

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FRUIT, a Scar-Free System for Targeted Chromosomal Mutagenesis, Epitope Tagging, and Promoter Replacement in Escherichia coli and Salmonella enterica 英文参考文献.doc

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FRUIT, a Scar-Free System for Targeted Chromosomal Mutagenesis, Epitope Tagging, and Promoter Replacement in Escherichia coli and Salmonella enterica 英文参考文献

FRUIT,aScar-FreeSystemforTargetedChromosomal Mutagenesis,EpitopeTagging,andPromoter ReplacementinEscherichiacoliandSalmonellaenterica AnneM.Stringer1,NavjotSingh1,AnastasiyaYermakova2,BriannaL.Petrone1, JayalekaJ.Amarasinghe1,LuciaReyes-Diaz3,NicholasJ.Mantis1,2,JosephT.Wade1,2 * 1Wadsworth Center,New York StateDepartment of Health, Albany, New York, United States ofAmerica, 2Department ofBiomedical Sciences, University at Albany, Albany,NewYork,UnitedStatesofAmerica,3UASRPCSTEPProgram,UniversityatAlbany,Albany,NewYork,UnitedStatesofAmerica Abstract Recombineering is a widely-used approach to delete genes, introduce insertions and point mutations, and introduce epitope tags into bacterial chromosomes. Many recombineering methods have been described, for a wide range of bacterial species. These methods are often limited by (i) low efficiency, and/or (ii) introduction of ‘‘scar’’ DNA into the chromosome.Here,wedescribearapid,efficient,PCR-basedrecombineeringmethod,FRUIT,thatcanbeusedtointroduce scar-freepointmutations,deletions,epitopetags, andpromotersintothegenomesofentericbacteria.Theefficiencyof FRUITisfarhigherthanthatofthemostwidely-usedrecombineeringmethodforEscherichiacoli.WehaveusedFRUITto introducepointmutationsandepitopetagsintothechromosomesofE.coliK-12,EnterotoxigenicE.coli,andSalmonella enterica.WehavealsousedFRUITtointroduceconstitutiveandinduciblepromotersintothechromosomeofE.coliK-12. Thus,FRUITisaversatile,efficientrecombineeringapproachthatcanbeappliedinmultiplespeciesofentericbacteria. Citation: Stringer AM, Singh N, Yermakova A, Petrone BL, Amarasinghe JJ, et al. (2012) FRUIT, a Scar-Free System for Targeted Chromosomal Mutagenesis, EpitopeTagging,andPromoterReplacementinEscherichiacoliandSalmonellaenterica.PLoSONE7(9):e44841.doi:10.1371/journal.pone.0044841 Editor:MarkAlexanderWebber,UniversityofBirmingham,UnitedKingdom ReceivedJune5,2012;AcceptedAugust7,2012;PublishedSeptember27,2012 Copyright: ? 2012 Stringer et al. This is an open-access art