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In Vitro and In Vivo Methodologies for Studying the Sigma 54-Dependent Transcription

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Bacterial Transcriptional Control

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1276))

Abstract

Here we describe approaches and methods to assaying in vitro the major variant bacterial sigma factor, Sigma 54 (σ54), in a purified system. We include the complete transcription system, binding interactions between σ54 and its activators, as well as the self-assembly and the critical ATPase activity of the cognate activators which serve to remodel the closed promoter complexes. We also present in vivo methodologies that are used to study the impact of physiological processes, metabolic states, global signalling networks, and cellular architecture on the control of σ54-dependent gene expression.

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Acknowledgements

This work was supported by BBSRC (BB/J002828/1), Wellcome Trust (WT093044MA), and Leverhulme Trust (F/07 058/BM) project grants.

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Authors and Affiliations

  1. Department of Life Sciences, Imperial College London, South Kensington Campus, London, SW7 2AZ, UK

    Martin Buck, Christoph Engl, Nicolas Joly, Goran Jovanovic, Milija Jovanovic, Edward Lawton, Christopher McDonald, Jörg Schumacher, Christopher Waite & Nan Zhang

  2. Equipe Nanomanipulation de Biomolecules, Institut Jacques Monod – UMR7592, CNRS – Universite Paris, Paris Cedex 13, 75205, France

    Nicolas Joly

Authors
  1. Martin Buck
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  2. Christoph Engl
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  3. Nicolas Joly
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  4. Goran Jovanovic
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  5. Milija Jovanovic
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  6. Edward Lawton
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  7. Christopher McDonald
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  8. Jörg Schumacher
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  9. Christopher Waite
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  10. Nan Zhang
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Corresponding author

Correspondence to Martin Buck .

Editor information

Editors and Affiliations

  1. Department of Microbiology, The Ohio State University, Columbus, Ohio, USA

    Irina Artsimovitch

  2. Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, Colorado, USA

    Thomas J. Santangelo

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Buck, M. et al. (2015). In Vitro and In Vivo Methodologies for Studying the Sigma 54-Dependent Transcription. In: Artsimovitch, I., Santangelo, T. (eds) Bacterial Transcriptional Control. Methods in Molecular Biology, vol 1276. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2392-2_4

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  • DOI: https://doi.org/10.1007/978-1-4939-2392-2_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2391-5

  • Online ISBN: 978-1-4939-2392-2

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