RMDAP A Versatile, Ready-To-Use Toolbox for Multigene Genetic Transformation 英文参考文献.docVIP

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RMDAP A Versatile, Ready-To-Use Toolbox for Multigene Genetic Transformation 英文参考文献.doc

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RMDAP A Versatile, Ready-To-Use Toolbox for Multigene Genetic Transformation 英文参考文献

RMDAP:AVersatile,Ready-To-UseToolboxforMultigene GeneticTransformation LeiMa,JiangliDong,YongshengJin,MingliangChen,XiaoyeShen,TaoWang* StateKeyLaboratoryofAgrobiotechnology,CollegeofBiologicalSciences,ChinaAgriculturalUniversity,Beijing,China Abstract Background: The use of transgenes to improve complex traits in crops has challenged current genetic transformation technologyformultigenetransfer.Therefore,amultigenetransformationstrategyforuseinplantmolecularbiologyand plantgeneticbreedingisthusneeded. Methodology/Principal Findings: Here we describe a versatile, ready-to-use multigene genetic transformation method, namedtheRecombination-assistedMultifunctionalDNAAssemblyPlatform(RMDAP),whichcombinesmanyoftheuseful features of existing plant transformation systems. This platform incorporates three widely-used recombination systems, namely,Gatewaytechnology,invivoCre/loxPandrecombineeringintoahighlyefficientandreliableapproachforgene assembly.RMDAPproposesastrategyforgenestackingandcontainsawiderangeofflexible,modularvectorsofferinga seriesoffunctionallyvalidatedgeneticelementstomanipulatetransgeneoverexpressionorgenesilencinginvolvedina metabolicpathway.Inparticular,theabilitytoconstructamultigenemarker-freevectorisanotherattractivefeature.The built-in flexibility of original vectors has greatly increased the expansibility and applicability of the system. A proof-of- principleexperimentwasconfirmedbysuccessfullytransferringseveralheterologousgenesintotheplantgenome. Conclusions/Significance: This platform is a ready-to-use toolbox for full exploitation of the potential for coordinate regulationofmetabolicpathwaysandmolecularbreeding,andwilleventuallyachievetheaimofwhatwecall‘‘one-stop breeding.’’ Citation:MaL,DongJ,JinY,ChenM,ShenX,etal.(2011)RMDAP:AVersatile,Ready-To-UseToolboxforMultigeneGeneticTransformation.PLoSONE6(5): e19883.doi:10.1371/journal.pone.0019883 Editor:HanyA.El-Shemy,CairoUniversity,Egypt ReceivedAugust16,2010;AcceptedApril20,2011;PublishedMay13,