Rational Design of a Plasmid Origin That Replicates Efficiently in Both Gram-Positive and Gram-Negative Bacteria 英文参考文献.docVIP

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Rational Design of a Plasmid Origin That Replicates Efficiently in Both Gram-Positive and Gram-Negative Bacteria 英文参考文献.doc

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Rational Design of a Plasmid Origin That Replicates Efficiently in Both Gram-Positive and Gram-Negative Bacteria 英文参考文献

RationalDesignofaPlasmidOriginThatReplicates EfficientlyinBothGram-PositiveandGram-Negative Bacteria AntonV.Bryksin,IchiroMatsumura* CenterforFundamentalandAppliedMolecularEvolution,DepartmentofBiochemistry,EmoryUniversity,Atlanta,Georgia,UnitedStatesofAmerica Abstract Background:Mostplasmidsreplicateonlywithinaparticulargenusorfamily. Methodology/PrincipalFindings:Herewedescribeanengineeredhighcopynumberexpressionvector,pBAV1K-T5,that produces varying quantities of active reporter proteins in Escherichia coli, Acinetobacter baylyi ADP1, Agrobacterium tumefaciens, (all Gram-negative), Streptococcus pneumoniae, Leifsonia shinshuensis, Peanibacillus sp. S18-36 and Bacillus subtilis(Gram-positive). Conclusions/Significance: Our results demonstrate the efficiency of pBAV1K-T5 replication in different bacterial species, therebyfacilitatingthestudyofproteinsthatdon’tfoldwellinE.coliandpathogensnotamenabletoexistinggenetictools. Citation:BryksinAV,MatsumuraI(2010)RationalDesignofaPlasmidOriginThatReplicatesEfficientlyinBothGram-PositiveandGram-NegativeBacteria.PLoS ONE5(10):e13244.doi:10.1371/journal.pone.0013244 Editor:IgorMokrousov,St.PetersburgPasteurInstitute,RussianFederation ReceivedJuly25,2010;AcceptedSeptember8,2010;PublishedOctober8,2010 Copyright:?2010Bryksin,Matsumura.Thisisanopen-accessarticledistributedunderthetermsoftheCreativeCommonsAttributionLicense,whichpermits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:TheworkwassupportedbytheNationalInstitutesofHealthgrants1R01GM074264and1R01GM086824toIM.Thefunderhadnoroleinstudy design,datacollectionandanalysis,decisiontopublish,orpreparationofthemanuscript. CompetingInterests:Theauthorshavedeclaredthatnocompetinginterestsexist. *E-mail:imatsum@ Introduction protein remains bound to the 59 phosphate after the nicking action.Thenewlyreleased39hydroxylontheoppositeendserves asaprimerforDNAsynthesis.ThehostDNApolymeraseusesthe unnickedcircularstrandas