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In Vivo DNA Analysis

  • Protocol
Book cover DNA-Protein Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 148))

Abstract

The in vivo analysis of DNA-protein interactions and chromatin structure can provide several kinds of critical information regarding regulation of gene expression and gene function. For example, DNA sequences spanned by nuclease-hypersensitive sites or bound by transcription factors often correspond to genetic regulatory elements. Using the ligation-mediated polymerase chain reaction (LMPCR) technology it is possible to map such DNA sequences and to demonstrate the existence of unusual DNA structures directly in living cells. LMPCR analyses can thus be used as a primary investigative tool to identify the regulatory sequences involved in gene expression. Once specific promoter sequence sites are shown to be bound by transcription factors in living cells, it is often possible to establish the identity of these factors simply by comparison with the consensus binding sites of known factors such as Sp1, AP-1, NF-1, and so forth. The identity of each factor can then be confirmed using in vitro gel shift (electrophoretic mobility shift assay [EMSA]) or footprinting assays.

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Author information

Authors and Affiliations

  1. Department de Biologie Médicale, Université Laval, et Unité de Recherche en Génétique Humaine et Moléculaire, Centre de Recherche, Pavilion Saint-Francois d’Assise, Québec, Canada

    Régen Drouin, Jean-Philippe Therrien, Martin Angers & Stéphane Ouellet

Authors
  1. Régen Drouin
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  2. Jean-Philippe Therrien
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  3. Martin Angers
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  4. Stéphane Ouellet
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Editors and Affiliations

  1. Centre de Recherche en Cancérologie de l’Université Laval, Centre Hopital Universitaire de Québec et Départment de biologie médicale, Université Laval, Québec, QC, Canada

    Tom Moss

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© 2001 Humana Press Inc., Totowa, NJ

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Drouin, R., Therrien, JP., Angers, M., Ouellet, S. (2001). In Vivo DNA Analysis. In: Moss, T. (eds) DNA-Protein Interactions. Methods in Molecular Biology, vol 148. Humana Press. https://doi.org/10.1385/1-59259-208-2:175

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  • DOI: https://doi.org/10.1385/1-59259-208-2:175

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-625-3

  • Online ISBN: 978-1-59259-208-1

  • eBook Packages: Springer Protocols

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