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dc.contributor.authorCebrián Castillo, Rubénes_ES
dc.contributor.authorRodríguez Ruano, Sonia es_ES
dc.contributor.authorMartínez Bueno, Manuel es_ES
dc.contributor.authorValdivia Martínez, Dolores Eva es_ES
dc.contributor.authorMaqueda Abreu, Mercedes es_ES
dc.contributor.authorMontalbán-López, Manueles_ES
dc.date.accessioned2014-03-25T07:55:18Z
dc.date.available2014-03-25T07:55:18Z
dc.date.issued2014
dc.identifier.citationCebrián, R.; et al. Analysis of the Promoters Involved in Enterocin AS-48 Expression. Plos One, 9(3): e90603 (2014). [http://hdl.handle.net/10481/31075]es_ES
dc.identifier.issn1932-6203
dc.identifier.otherdoi: 10.1371/journal.pone.0090603
dc.identifier.urihttp://hdl.handle.net/10481/31075
dc.description.abstractThe enterocin AS-48 is the best characterized antibacterial circular protein in prokaryotes. It is a hydrophobic and cationic bacteriocin, which is ribosomally synthesized by enterococcal cells and post-translationally cyclized by a head-to-tail peptide bond. The production of and immunity towards AS-48 depend upon the coordinated expression of ten genes organized in two operons, as-48ABC (where genes encoding enzymes with processing, secretion, and immunity functions are adjacent to the structural as-48A gene) and as-48C1DD1EFGH. The current study describes the identification of the promoters involved in AS-48 expression. Seven putative promoters have been here amplified, and separately inserted into the promoter-probe vector pTLR1, to create transcriptional fusions with the mCherry gene used as a reporter. The activity of these promoter regions was assessed measuring the expression of the fluorescent mCherry protein using the constitutive pneumococcal promoter PX as a reference. Our results revealed that only three promoters PA, P2(2) and PD1 were recognized in Enterococcus faecalis, Lactococcus lactis and Escherichia coli, in the conditions tested. The maximal fluorescence was obtained with PX in all the strains, followed by the P2(2) promoter, which level of fluorescence was 2-fold compared to PA and 4-fold compared to PD1. Analysis of putative factors influencing the promoter activity in single and double transformants in E. faecalis JH2-2 demonstrated that, in general, a better expression was achieved in presence of pAM401-81. In addition, the P2(2) promoter could be regulated in a negative fashion by genes existing in the native pMB-2 plasmid other than those of the as-48 cluster, while the pH seems to affect differently the as-48 promoter expression.es_ES
dc.description.sponsorshipThis work was supported in part by the Ministerio de Ciencia e Innovación project BIO2008-01708, the Plan Propio from the University of Granada (Spain) and by the Research Plan Group (BIO 160).es_ES
dc.language.isoenges_ES
dc.publisherPublic Library of Science (PLOS)es_ES
dc.rightsCreative Commons Attribution-NonCommercial-NoDerivs 3.0 License
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/3.0/
dc.subjectDNA transcriptionen_EN
dc.subjectEnterococcus faecalisen_EN
dc.subjectFluorescence en_EN
dc.subjectGene expressionen_EN
dc.subjectLactococcus lactisen_EN
dc.subjectOperonsen_EN
dc.subjectPromoter regionsen_EN
dc.subjectSequence motif analysisen_EN
dc.titleAnalysis of the Promoters Involved in Enterocin AS-48 Expressionen_EN
dc.typeinfo:eu-repo/semantics/articlees_ES
dc.rights.accessRightsinfo:eu-repo/semantics/openAccesses_ES


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