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Engineering Green-light-responsive Heterologous Gene Expression in Pseudomonas

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Pseudomonas aeruginosa

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2721))

Abstract

Engineering bacterial properties requires precision and fine-tuning for optimal control of the desired application. In consequence, it is essential to accurately turn the function of interest from OFF to ON state and vice versa, avoiding any type of residual activation. For this type of purpose, light switches have revealed a clean and powerful tool in which control does not depend on the addition of chemical compounds that may remain in the media. To reach this degree of directed regulation through light, the switch based on the cyanobacterial two-component system CcaSR system was previously adapted to manipulate Pseudomonas putida for transcription of a gene of interest. In this chapter, we describe how to induce biofilm formation by placing the expression of the c-di-GMP-producing diguanylate cyclase PleD from Caulobacter sp. under the control of the CcaSR system. The regulation through optogenetics accomplished with this protocol promotes higher exploitation of biofilm beneficial features in a cheaper and cleaner way compared to chemical induction.

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Acknowledgments

This work was funded by the SETH (RTI2018-095584-B-C42) (MINECO/FEDER) and SyCoLiM (ERA-COBIOTECH 2018 - PCI2019-111859-2) Projects of the Spanish Ministry of Science and Innovation, the MADONNA (H2020-FET-OPEN-RIA-2017-1-766975), BioRoboost (H2020-NMBP-BIO-CSA-2018-820699), SynBio4Flav (H2020-NMBP-TR-IND/H2020-NMBP-BIO-2018-814650) and MIX-UP (MIX-UP H2020-BIO-CN-2019-870294) Contracts of the European Union and the InGEMICS-CM (S2017/BMD-3691) Project of the Comunidad de Madrid - European Structural and Investment Funds (FSE, FECER).

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Authors and Affiliations

  1. Systems Biology Department, Centro Nacional de Biotecnología (CNB-CSIC), Madrid, Spain

    Angeles Hueso-Gil, Belén Calles & Víctor de Lorenzo

  2. Centro de Biotecnología y Genómica de Plantas (UPM-INIA/CSIC), Madrid, Spain

    Angeles Hueso-Gil

Authors
  1. Angeles Hueso-Gil
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  2. Belén Calles
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  3. Víctor de Lorenzo
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Corresponding author

Correspondence to Víctor de Lorenzo .

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Editors and Affiliations

  1. Department of Biosciences, Università degli Studi di Milano, Milan, Milano, Italy

    Giovanni Bertoni

  2. Department of Biosciences, Università degli Studi di Milano, Milan, Milano, Italy

    Silvia Ferrara

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Hueso-Gil, A., Calles, B., de Lorenzo, V. (2024). Engineering Green-light-responsive Heterologous Gene Expression in Pseudomonas. In: Bertoni, G., Ferrara, S. (eds) Pseudomonas aeruginosa. Methods in Molecular Biology, vol 2721. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3473-8_3

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  • DOI: https://doi.org/10.1007/978-1-0716-3473-8_3

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3472-1

  • Online ISBN: 978-1-0716-3473-8

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