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dc.contributor.authorRomero-Jiménez, Lorena-
dc.contributor.authorRodríguez-Carbonell, David-
dc.contributor.authorGallegos, María Trinidad-
dc.contributor.authorSanjuán, Juan-
dc.contributor.authorPérez-Mendoza, Daniel-
dc.date.accessioned2015-11-25T17:57:10Z-
dc.date.available2015-11-25T17:57:10Z-
dc.date.issued2015-09-29-
dc.identifier.citationBMC Microbiology 15(1): 190 (2015)-
dc.identifier.issn1471-2180-
dc.identifier.urihttp://hdl.handle.net/10261/125916-
dc.description.abstract[Background] The cyclic diguanylate (c-di-GMP) is currently considered an ubiquitous second messenger in bacteria that influences a wide range of cellular processes. One of the methodological approaches to unravel c-di-GMP regulatory networks involves raising the c-di-GMP intracellular levels, e.g. by expressing a diguanylate cyclase (DGC), to provoke phenotypic changes.-
dc.description.abstract[Results] We have constructed mini-Tn7 delivery vectors for the integration and stable expression of the pleD* gene encoding a highly active DGC, which can be used to artificially increase the intracellular levels of c-di-GMP in Gram negative bacteria. The functionality of these new vectors has been validated in several plant-interacting α- and γ-proteobacteria. Similarly to vector plasmid-borne pleD*, the genome-borne mini-Tn7pleD* constructs provide significant increases in intracellular c-di-GMP, provoking expected phenotypic changes such as enhanced polysaccharide production, biofilm formation and reduced motility. However, the mini-Tn7pleD* constructs resulted far more stable in the absence of antibiotics than the plasmid-based pleD* constructs. Furthermore, we have also implemented an inducible system to modulate pleD* expression and intracellular c-di-GMP rises “on demand”.-
dc.description.abstract[Conclusions] mini-Tn7pleD* constructs are very stable and are maintained during bacterial free-living growth as well as during interaction with eukaryotic hosts, in the absence of selective pressure. This high stability ensures experimental homogeneity in time and space with regard to enhancing c-di-GMP intracellular levels in bacteria of interest.-
dc.description.sponsorshipThis work was supported by grants BIO2011-23032 and BIO2014-55075-P (Ministerio de Economía y Competitividad) and P10-CVI-5800 (Junta de Andalucía), all co-financed with FEDER funds, and CSIC 201440E026. LRJ was supported by JAE-Pre fellowship, and DRC by a contract associated to BIO2011-23032. DPM was supported by a JAE-Doc grant and contracts associated to grants P10-CVI-5800 and CSIC 201440E026.-
dc.publisherBioMed Central-
dc.relation.isversionofPublisher's version-
dc.rightsopenAccess-
dc.subjectc-di-GMP-
dc.subjectMini-Tn7-
dc.subjectSignal transduction-
dc.subjectBiofilms-
dc.subjectExopolysaccharide production-
dc.subjectBacterial motility-
dc.subjectPlasmid stability-
dc.titleMini-Tn7 vectors for stable expression of diguanylate cyclase PleD* in Gram-negative bacteria-
dc.typeArtículo-
dc.identifier.doi10.1186/s12866-015-0521-6-
dc.relation.publisherversionhttp://dx.doi.org/10.1186/s12866-015-0521-6-
dc.date.updated2015-11-25T17:57:10Z-
dc.language.rfc3066en-
dc.rights.holderRomero-Jiménez et al.-
dc.rights.licensehttp://creativecommons.org/licenses/by/4.0/-
dc.contributor.funderMinisterio de Economía y Competitividad (España)-
dc.contributor.funderJunta de Andalucía-
dc.contributor.funderEuropean Commission-
dc.contributor.funderConsejo Superior de Investigaciones Científicas (España)-
dc.relation.csic-
dc.identifier.funderhttp://dx.doi.org/10.13039/501100003329es_ES
dc.identifier.funderhttp://dx.doi.org/10.13039/501100000780es_ES
dc.identifier.funderhttp://dx.doi.org/10.13039/501100003339es_ES
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