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Fluorescent protein vectors for promoter analysis in lactic acid bacteria and Escherichia coli

  • Applied genetics and molecular biotechnology
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Abstract

Fluorescent reporter genes are valuable tools for real-time monitoring of gene expression in living cells. In this study we describe the construction of novel promoter-probe vectors containing a synthetic mCherry fluorescent protein gene, codon-optimized for lactic acid bacteria, divergently linked, or not, to a gene encoding the S65T and F64L variant of the green fluorescent protein. The utility of the transcriptional fusion vectors was demonstrated by the cloning of a single or two divergent promoter regions and by the quantitative evaluation of fluorescence during growth of Lactococcus lactis, Enterococcus faecalis, and Escherichia coli.

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Acknowledgments

This work was supported by the Spanish Ministry of Science and Innovation (grants AGL2009-12998-C03-01, AGL2009-13361-C02-02, and CSD2007-00063 Consolider Ingenio 2010 FUN-C-FOOD), the Comunidad de Madrid (grant ALIBIRD P2009/AGR-1469), and the European Union VII framework program (Initial Training Network, grant no. 238490).

We thank Dr. Stephen Elson for the critical reading of the manuscript. We are grateful to M. Angeles Corrales for her technical assistance.

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Authors and Affiliations

  1. Departamento de Biotecnología y Microbiología de Alimentos, Instituto de Investigación en Ciencias de la Alimentación CIAL (CSIC-UAM), CEI UAM+CSIC, Nicolás Cabrera 9, 28049, Madrid, Spain

    Tomás García-Cayuela, Luz P. Gómez de Cadiñanos, Carmen Peláez & Teresa Requena

  2. Departamento de Microbiología Molecular y Biología de las Infecciones, Centro de Investigaciones Biológicas CIB (CSIC), Madrid, Spain

    M. Luz Mohedano, Pilar Fernández de Palencia & Paloma López

  3. Janssen Pharmaceutica, Beerse, Belgium

    Daniel Boden

  4. Host-Microbe Interactomics, University of Wageningen, Wageningen, The Netherlands

    Jerry Wells

Authors
  1. Tomás García-Cayuela
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  2. Luz P. Gómez de Cadiñanos
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  3. M. Luz Mohedano
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  4. Pilar Fernández de Palencia
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  5. Daniel Boden
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  6. Jerry Wells
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  7. Carmen Peláez
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  8. Paloma López
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  9. Teresa Requena
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Corresponding author

Correspondence to Teresa Requena.

Additional information

Tomás García-Cayuela, Luz P. Gómez de Cadiñanos, and M. Luz Mohedano contributed equally to this work.

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García-Cayuela, T., de Cadiñanos, L.P.G., Mohedano, M.L. et al. Fluorescent protein vectors for promoter analysis in lactic acid bacteria and Escherichia coli . Appl Microbiol Biotechnol 96, 171–181 (2012). https://doi.org/10.1007/s00253-012-4087-z

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  • DOI: https://doi.org/10.1007/s00253-012-4087-z

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