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Título: | Novel plasmid-based genetic tools for the study of promoters and terminators in Streptococcus pneumoniae and Enterococcus faecalis. |
Autor: | Ruiz-Cruz, Sofía CSIC ORCID; Solano-Collado, Virtu CSIC ORCID; Espinosa, Manuel CSIC ORCID ; Bravo, Alicia CSIC ORCID | Palabras clave: | Gram-positive bacteria Enterococcus faecalis Streptococcus agalactiae Streptococcus pneumoniae Probe plasmid vectors pMV158 Genetic promoters Genetic terminators Green fluorescent proteins |
Fecha de publicación: | nov-2010 | Editor: | Elsevier | Citación: | Journal of Microbiological Methods, 83(2): 156-163 (2010) | Resumen: | Promoter-probe and terminator-probe plasmid vectors make possible to rapidly examine whether particular sequences function as promoter or terminator signals in various genetic backgrounds and under diverse environmental stimuli. At present, such plasmid-based genetic tools are very scarce in the Gram-positive pathogenic bacteria Streptococcus pneumoniae and Enterococcus faecalis. Hence, we developed novel promoter-probe and terminator-probe vectors based on the Streptococcus agalactiae pMV158 plasmid, which replicates autonomously in numerous Gram-positive bacteria. As reporter gene, a gfp allele encoding a variant of the green fluorescent protein was used. These genetic tools were shown to be suitable to assess the activity of promoters and terminators (both homologous and heterologous) in S. pneumoniae and E. faecalis. In addition, the promoter-probe vector was shown to be a valuable tool for the analysis of regulated promoters in vivo, such as the promoter of the pneumococcal fuculose kinase gene. These new plasmid vectors will be very useful for the experimental verification of predicted promoter and terminator sequences, as well as for the construction of new inducible-expression vectors. Given the promiscuity exhibited by the pMV158 replicon, these vectors could be used in a variety of Gram-positive bacteria. | Descripción: | Este artículo pertenece a la Tesis presentada por Virtudes Solano Collado con título: "Caracterización molecular del regulador transcripcional MgaSpn de Streptococcus pneumoniae"(https://digital.csic.es/handle/10261/98162) | Versión del editor: | http://dx.doi.org/10.1016/j.mimet.2010.08.004 | URI: | http://hdl.handle.net/10261/42052 | DOI: | 10.1016/j.mimet.2010.08.004 | ISSN: | 1872-8359 |
Aparece en las colecciones: | (CIB) Artículos |
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Plasmids-Ruiz-Cruz et al.pdf | 643,92 kB | Adobe PDF | Visualizar/Abrir |
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