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Bacterial Artificial Chromosome Mutagenesis Using Recombineering英文文献资料.doc

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Bacterial Artificial Chromosome Mutagenesis Using Recombineering英文文献资料

HindawiPublishingCorporation JournalofBiomedicineandBiotechnology Volume2011,ArticleID971296,10pages doi:10.1155/2011/971296 ReviewArticle BacterialArti?cialChromosomeMutagenesisUsing Recombineering KumaranNarayanan1,2andQingwenChen 2 1 2 DepartmentofGeneticsandGenomicSciences,MountSinaiSchoolofMedicine,NewYork,NY10029,USA SchoolofScience,MonashUniversity,SunwayCampus,Room2-5-29,BandarSunway,46150,Malaysia CorrespondenceshouldbeaddressedtoKumaranNarayanan,kumaran.narayanan@.my Received2August2010;Accepted21October2010 AcademicEditor:MasamitsuYamaguchi Copyright?2011K.NarayananandQ.Chen.ThisisanopenaccessarticledistributedundertheCreativeCommonsAttribution License,whichpermitsunrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalworkisproperly cited. Geneexpressionfrombacterialarti?cialchromosome(BAC)cloneshasbeendemonstratedtofacilitatephysiologicallyrelevant levelscomparedtoviralandnonviralcDNAvectors.BACsarelargeenoughtotransferintactgenesintheirnativechromosomal settingtogetherwith ?ankingregulatoryelementstoprovideallthesignalsforcorrectspatiotemporalgeneexpression.Until recently, the use of BACs for functional studies has been limited because their large size has inherently presented a major obstacleforintroducingmodi?cationsusingconventionalgeneticengineeringstrategies.Thedevelopmentofinvivohomologous recombinationstrategiesbasedonrecombineeringinE.colihashelpedresolvethisproblembyenablingfacileengineeringofhigh molecularweightBACDNAwithoutdependenceonsuitablyplacedrestrictionenzymesorcloningsteps.Thesetechniqueshave considerablyexpandedthepossibilitiesforstudyingfunctionalgeneticsusingBACsinvitroandinvivo. 1.Introduction an attractive resource for studying genome structure and function,representinganexcitingalternativetoconventional vectorsystems. TheinformationgeneratedfromtheGenomeProjectswill beofthegreatestvalueifitcanbeconvertedintofunctional data, particularly if this increases our understanding of normalgenefunctionandallowsstra