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dc.contributor.authorMarx, Christopher J
dc.date.accessioned2010-10-05T15:36:29Z
dc.date.issued2008
dc.identifier.citationMarx, Christopher J. 2008. Development of a broad-host-range sacB-based vector for unmarked allelic exchange. BMC Research Notes 1:1.en_US
dc.identifier.issn1756-0500en_US
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:4457689
dc.description.abstractBackground: Although genome sequences are available for an ever-increasing number of bacterial species, the availability of facile genetic tools for physiological analysis have generally lagged substantially behind traditional genetic models. Results: Here I describe the development of an improved, broad-host-range "in-out" allelic exchange vector, pCM433, which permits the generation of clean, marker-free genetic manipulations. Wild-type and mutant alleles were reciprocally exchanged at three loci in Methylobacterium extorquens AM1 in order to demonstrate the utility of pCM433. Conclusion: The broad-host-range vector for marker-free allelic exchange described here, pCM433, has the advantages of a high copy, general Escherichia coli replicon for easy cloning, an IncP oriT enabling conjugal transfer, an extensive set of restriction sites in its polylinker, three antibiotic markers, and sacB (encoding levansucrase) for negative selection upon sucrose plates. These traits should permit pCM433 to be broadly applied across many bacterial taxa for marker-free allelic exchange, which is particularly important if multiple manipulations or more subtle genetic manipulations such as point mutations are desired.en_US
dc.description.sponsorshipOrganismic and Evolutionary Biologyen_US
dc.language.isoen_USen_US
dc.publisherBioMed Centralen_US
dc.relation.isversionofdoi:10.1186/1756-0500-1-1en_US
dc.relation.hasversionhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC2518277/pdf/en_US
dash.licenseOAP
dc.titleDevelopment of a Broad-Host-Range SacB-Based Vector for Unmarked Allelic Exchangeen_US
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden_US
dc.relation.journalBMC Research Notesen_US
dash.depositing.authorMarx, Christopher J
dc.date.available2010-10-05T15:36:29Z
dc.identifier.doi10.1186/1756-0500-1-1*
dash.contributor.affiliatedMarx, Christopher J


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