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Enrichment of Golgi Membranes from Triticum aestivum (Wheat) Seedlings

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Isolation of Plant Organelles and Structures

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1511))

Abstract

The Golgi apparatus is an essential component in the plant secretory pathway. The enrichment of Golgi membranes from plant tissue is fundamental to the study of this structurally complex organelle. The utilization of density centrifugation for the enrichment of Golgi membranes is still the most widely employed isolation technique. Generally, the procedure requires optimization depending on the plant tissue being employed. Here we provide a detailed enrichment procedure that has previously been used to characterize cell wall biosynthetic complexes from wheat seedlings. We also outline several downstream analyses procedures, including nucleoside diphosphatase assays, immunoblotting, and finally localization of putative Golgi proteins by fluorescent tags.

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Acknowledgments

This work was funded by grants from the Australia Research Council (ARC) to the ARC Centre of Excellence in Plant Cell Walls [CE110001007] and the U. S. Department of Energy, Office of Science, Office of Biological and Environmental Research, through contract DE-AC02-05CH11231 between Lawrence Berkeley National Laboratory and the U. S. Department of Energy. JLH is supported by an ARC Future Fellowship [FT130101165].

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Authors and Affiliations

  1. The ARC Centre of Excellence in Plant Cell Walls, School of Biosciences, The University of Melbourne, Melbourne, VIC, Australia

    Wei Zeng, Berit Ebert, Carsten Rautengarten & Antony Bacic

  2. Department of Biochemistry, University of Cambridge, Cambridge CB2 1QR, UK

    Berit Ebert & Carsten Rautengarten

  3. Joint BioEnergy Institute and Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94702, USA

    Harriet T. Parsons & Joshua L. Heazlewood

  4. School of BioSciences, The University of Melbourne, Melbourne, 3010, VIC, Australia

    Joshua L. Heazlewood

Authors
  1. Wei Zeng
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  2. Berit Ebert
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  3. Harriet T. Parsons
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  4. Carsten Rautengarten
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  5. Antony Bacic
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  6. Joshua L. Heazlewood
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Corresponding author

Correspondence to Joshua L. Heazlewood .

Editor information

Editors and Affiliations

  1. The University of Western Australia, Perth, West Australia, Australia

    Nicolas L. Taylor

  2. The University of Western Australia, Perth, West Australia, Australia

    A. Harvey Millar

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Zeng, W., Ebert, B., Parsons, H.T., Rautengarten, C., Bacic, A., Heazlewood, J.L. (2017). Enrichment of Golgi Membranes from Triticum aestivum (Wheat) Seedlings. In: Taylor, N., Millar, A. (eds) Isolation of Plant Organelles and Structures. Methods in Molecular Biology, vol 1511. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6533-5_11

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  • DOI: https://doi.org/10.1007/978-1-4939-6533-5_11

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6531-1

  • Online ISBN: 978-1-4939-6533-5

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