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Fluorescence-Activated Nucleolus Sorting in Arabidopsis

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Book cover The Nucleolus

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1455))

Abstract

Nucleolar isolation allows exhaustive characterization of the nucleolar content. Centrifugation-based protocols are not adapted to isolation of nucleoli directly from a plant tissue because of copurification of cellular debris. We describe here a method that allows the purification of nucleoli using fluorescent-activated cell sorting from Arabidopsis thaliana leaves. This approach requires the expression of a specific nucleolar protein such as fibrillarin fused to green fluorescent protein in planta.

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Acknowledgments

This work was supported by the CNRS, the UPVD foundation BQR 2014 FANoS and the ANR NucleoReg ANR-15-CE12-0013-01 to FP, and ANR SubCellif to JSV. The cell sorter is an instrument of the Montpellier Rio Imaging platform. The authors thank Richard Cooke for English corrections.

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Authors and Affiliations

  1. CNRS, Laboratoire Génome et Développement des Plantes, UMR5096, 58 Avenue P. Alduy, 66860, Perpignan, France

    Frédéric Pontvianne & Julio Sáez-Vásquez

  2. University Perpignan Via Domitia, Laboratoire Génome et Développement des Plantes, UMR5096, 66860, Perpignan, France

    Frédéric Pontvianne & Julio Sáez-Vásquez

  3. UMR5535, Institut de Génétique Moléculaire de Montpellier, 1919 route de Mende, 34293, Montpellier cedex 5, France

    Myriam Boyer-Clavel

  4. MRI UMS3426, BioCampus Montpellier, c/o IGF, 141 rue de la cardonille, 34094, Montpellier cedex 5, France

    Myriam Boyer-Clavel

Authors
  1. Frédéric Pontvianne
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  2. Myriam Boyer-Clavel
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  3. Julio Sáez-Vásquez
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Corresponding author

Correspondence to Frédéric Pontvianne .

Editor information

Editors and Affiliations

  1. Department of Biochemistry III, University of Regensburg, Regensburg, Germany

    Attila Németh

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© 2016 Springer Science+Business Media New York

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Pontvianne, F., Boyer-Clavel, M., Sáez-Vásquez, J. (2016). Fluorescence-Activated Nucleolus Sorting in Arabidopsis. In: Németh, A. (eds) The Nucleolus. Methods in Molecular Biology, vol 1455. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3792-9_15

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  • DOI: https://doi.org/10.1007/978-1-4939-3792-9_15

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3790-5

  • Online ISBN: 978-1-4939-3792-9

  • eBook Packages: Springer Protocols

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