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Companion Protease Inhibitors for the In Situ Protection of Recombinant Proteins in Plants

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Book cover Recombinant Proteins from Plants

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1385))

Abstract

We previously described a procedure for the use of plant protease inhibitors as “companion” accessory proteins to prevent unwanted proteolysis of clinically useful recombinant proteins in leaf crude protein extracts (Benchabane et al. Methods Mol Biol 483:265–273, 2009). Here we describe the use of these inhibitors for the protection of recombinant proteins in planta, before their extraction from leaf tissues. A procedure is first described involving inhibitors co-expressed along—and co-migrating—with the protein of interest in host plant cells. An alternative, single transgene scheme is then described involving translational fusions of the recombinant protein and companion inhibitor. These approaches may allow for a significant improvement of protein steady-state levels in leaves, comparable to yield improvements observed with protease-deficient strains of less complex protein expression hosts such as E. coli or yeasts.

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Acknowledgments

This work was supported by a Discovery grant from the Natural Science and Engineering Research Council (NSERC) of Canada to DM, and by an NSERC Strategic grant to DM funded in part by Medicago, Inc. P. Varennes-Jutras was the recipient of an NSERC/FRQ-NT BMP graduate scholarship supported by Medicago, Inc.

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Authors and Affiliations

  1. Centre de Recherche et d’Innovation sur les Végétaux, Université Laval, Québec, QC, Canada

    Stéphanie Robert, Philippe V. Jutras, Moustafa Khalf, Marie-Claire Goulet, Frank Sainsbury & Dominique Michaud

  2. Medicago Inc., Québec, QC, Canada

    Marc-André D’Aoust

  3. Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia, QLD, Australia

    Frank Sainsbury

Authors
  1. Stéphanie Robert
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  2. Philippe V. Jutras
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  3. Moustafa Khalf
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  4. Marc-André D’Aoust
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  5. Marie-Claire Goulet
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  6. Frank Sainsbury
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  7. Dominique Michaud
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Corresponding author

Correspondence to Dominique Michaud .

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Editors and Affiliations

  1. Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, London, Ontario, Canada

    Jacqueline MacDonald

  2. Department of Biology, Western University, London, Ontario, Canada

    Igor Kolotilin

  3. London, Ontario, Canada

    Rima Menassa

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Robert, S. et al. (2016). Companion Protease Inhibitors for the In Situ Protection of Recombinant Proteins in Plants. In: MacDonald, J., Kolotilin, I., Menassa, R. (eds) Recombinant Proteins from Plants. Methods in Molecular Biology, vol 1385. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3289-4_8

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  • DOI: https://doi.org/10.1007/978-1-4939-3289-4_8

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3288-7

  • Online ISBN: 978-1-4939-3289-4

  • eBook Packages: Springer Protocols

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