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Poly(ADP-Ribose) Polymerase pp 395–413Cite as

Purification of Recombinant Human PARG and Activity Assays

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1608))

Abstract

The purification of Poly(ADP-ribose) glycohydrolase (PARG) from overexpressing bacteria Escherichia coli is described here to a fast and reproducible one chromatographic step protocol. After cell lysis, GST-PARG-fusion proteins from the crude extract are affinity purified by a Glutathione 4B Sepharose chromatographic step. The PARG proteins are then freed from their GST-fusion by overnight enzymatic cleavage using the preScission protease. As described in the protocol, more than 500 μg of highly active human PARG can be obtained from 1.5 L of E. coli culture.

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Author information

Authors and Affiliations

  1. Groupe Poly(ADP-ribosyl)ation et Intégrité du Génome, UMR7242 du CNRS, École Supérieure de Biotechnologie de Strasbourg, Parc d’innovation, Boulevard Sébastien Brant CS 10413, 67412, Illkirch Cedex, France

    Jean-Christophe Amé, Éléa Héberlé, Barbara Camuzeaux, Françoise Dantzer & Valérie Schreiber

Authors
  1. Jean-Christophe Amé
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  2. Éléa Héberlé
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  3. Barbara Camuzeaux
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  4. Françoise Dantzer
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  5. Valérie Schreiber
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Corresponding author

Correspondence to Jean-Christophe Amé .

Editor information

Editors and Affiliations

  1. Cancer Biology Program, Fox Chase Cancer Center, Philadelphia, Pennsylvania, USA

    Alexei V. Tulin

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Amé, JC., Héberlé, É., Camuzeaux, B., Dantzer, F., Schreiber, V. (2017). Purification of Recombinant Human PARG and Activity Assays. In: Tulin, A. (eds) Poly(ADP-Ribose) Polymerase. Methods in Molecular Biology, vol 1608. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6993-7_25

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  • DOI: https://doi.org/10.1007/978-1-4939-6993-7_25

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6992-0

  • Online ISBN: 978-1-4939-6993-7

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