Abstract
The structure and dynamics of Escherichia coli transcription elongation complex are now well documented. However, most of the studies have been conducted in vitro and frequently under artificial conditions that facilitate the biochemical characterization of the complex. Thus, little is known about relevance of these results for the regulatory aspects of transcription elongation inside the cell. Here, we describe the use of a single-strand-specific probe chloroacetaldehyde for in situ footprinting of E. coli elongation complex temporarily halted by a protein roadblock. The method provides valuable information on the dynamic features of transcriptionally engaged RNA polymerase within the cellular environment.
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Acknowledgements
We thank Rachel Mooney and Bob Landick for the RL1600 and RL1601 strains. The work described in this chapter was supported in part by la Ligue contre le Cancer Grand Ouest and recurrent funding from the CNRS.
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Rahmouni, A.R., Mosrin-Huaman, C. (2015). In Situ Footprinting of E. coli Transcription Elongation Complex with Chloroacetaldehyde. 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_13
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DOI: https://doi.org/10.1007/978-1-4939-2392-2_13
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Publisher Name: Humana Press, New York, NY
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