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dc.contributor.authorBlas, Emilio-
dc.contributor.authorCava, Felipe-
dc.contributor.authorLópez-Viñas, Eduardo-
dc.contributor.authorMendieta, Jesús-
dc.contributor.authorBerenguer, José-
dc.date.accessioned2008-10-21T14:42:55Z-
dc.date.available2008-10-21T14:42:55Z-
dc.date.issued2007-06-29-
dc.identifier.citationApplied and Environmental Microbiology, 2007, p. 5138-5145, Vol. 73, No. 16en_US
dc.identifier.issn0099-2240-
dc.identifier.urihttp://hdl.handle.net/10261/7926-
dc.descriptionSupplementary material availableen_US
dc.description.abstractA spontaneous rpsL mutant of Thermus thermophilus was isolated in a search for new selection markers for this organism. This new allele, named rpsL1, encodes a K47R/K57E double mutant S12 ribosomal protein that confers a streptomycin-dependent (SD) phenotype to T. thermophilus. Models built on the available three-dimensional structures of the 30S ribosomal subunit revealed that the K47R mutation directly affects the streptomycin binding site on S12, whereas the K57E does not apparently affect this binding site. Either of the two mutations conferred the SD phenotype individually. The presence of the rpsL1 allele, either as a single copy inserted into the chromosome as part of suicide plasmids or in multicopy as replicative plasmids, produced a dominant SD phenotype despite the presence of a wild-type rpsL gene in a host strain. This dominant character allowed us to use the rpsL1 allele not only for positive selection of plasmids to complement a kanamycin-resistant mutant strain, but also more specifically for the isolation of deletion mutants through a single step of negative selection on streptomycin-free growth mediumen_US
dc.description.sponsorshipThis work has been supported by grants of codes BIO2004-02671, cofunded by the Ministerio de Educación y Ciencia and the European Union, and S0505/PPQ/0344, from Comunidad Autónoma de Madrid. Financial support of Fundación Ramón Areces to CBMSO is acknowledgeden_US
dc.format.extent236706 bytes-
dc.format.extent1831806 bytes-
dc.format.mimetypeapplication/pdf-
dc.format.mimetypeimage/jpeg-
dc.language.isoengen_US
dc.publisherAmerican Society for Microbiologyen_US
dc.rightsopenAccessen_US
dc.subjectThermus thermophilusen_US
dc.titleUse of a Dominant rpsL Allele Conferring Streptomycin Dependence for Positive and Negative Selection in Thermus thermophilusen_US
dc.typeartículoen_US
dc.identifier.doihttp://dx.doi.org/10.1128/AEM.00751-07-
dc.description.peerreviewedPeer revieweden_US
dc.relation.publisherversionhttp://dx.doi.org/10.1128/AEM.00751-07en_US
dc.identifier.e-issn1098-5336-
dc.contributor.funderMinisterio de Educación y Ciencia (España)-
dc.contributor.funderEuropean Commission-
dc.contributor.funderComunidad de Madrid-
dc.contributor.funderFundación Ramón Areces-
dc.identifier.funderhttp://dx.doi.org/10.13039/501100000780es_ES
dc.identifier.funderhttp://dx.doi.org/10.13039/100008054es_ES
dc.identifier.funderhttp://dx.doi.org/10.13039/100012818es_ES
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