Engineering Clostridium Strain to Accept Unmethylated DNA 英文参考文献.docVIP

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Engineering Clostridium Strain to Accept Unmethylated DNA 英文参考文献.doc

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Engineering Clostridium Strain to Accept Unmethylated DNA 英文参考文献

EngineeringClostridiumStraintoAcceptUnmethylated DNA HongjunDong1,2,YanpingZhang1,ZongjieDai1,3,YinLi1* 1Institute of Microbiology, Chinese Academy of Sciences, Beijing, China, 2Graduate School of the Chinese Academy of Sciences, Beijing, China, 3Department of BiochemistryandMolecularBiology,UniversityofScienceandTechnologyofChina,Hefei,China Abstract It is difficult to genetically manipulate the medically and biotechnologically important genus Clostridium due to the existence of the restriction and modification (RM) systems. We identified and engineered the RM system of a model clostridialspecies,C.acetobutylicum,withtheaimtoallowthehosttoaccepttheunmethylatedDNAefficiently.Agene CAC1502 putatively encoding the type II restriction endonuclease Cac824I was identified from the genome of C. acetobutylicumDSM1731,anddisruptedusingtheClosTronsystembasedongroupIIintroninsertion.Theresultingstrain SMB009lostthetypeIIrestrictionendonucleaseactivity,andcanbetransformedwithunmethylatedDNAasefficientlyas with methylated DNA. The strategy reported here makes it easy to genetically modify the clostridial species using unmethylatedDNA,whichwillhelptoadvancetheunderstandingoftheclostridialphysiologyfromthemolecularlevel. Citation: Dong H, Zhang Y, Dai Z, Li Y (2010) Engineering Clostridium Strain to Accept Unmethylated DNA. PLoS ONE 5(2): e9038. doi:10.1371/journal. pone.0009038 Editor:JianR.Lu,TheUniversityofManchester,UnitedKingdom ReceivedSeptember29,2009;AcceptedJanuary17,2010;PublishedFebruary9,2010 Copyright: ? 2010 Dong et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding: Thisworkwassupported bytheNationalHighTechnologyResearchandDevelopment ProgramofChina(863Project, #2006AA02Z237),National BasicResearchProgramofChina(973Project,#2007CB707803),KnowledgeInnovationProgramofCAS(#KSCXZ