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Recombinant Proteins from Plants

Volume 1385 of the series Methods in Molecular Biology pp 115-126

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

  • Stéphanie RobertAffiliated withCentre de Recherche et d’Innovation sur les Végétaux, Université Laval
  • , Philippe V. JutrasAffiliated withCentre de Recherche et d’Innovation sur les Végétaux, Université Laval
  • , Moustafa KhalfAffiliated withCentre de Recherche et d’Innovation sur les Végétaux, Université Laval
  • , Marc-André D’AoustAffiliated withMedicago Inc.
  • , Marie-Claire GouletAffiliated withCentre de Recherche et d’Innovation sur les Végétaux, Université Laval
  • , Frank SainsburyAffiliated withCentre de Recherche et d’Innovation sur les Végétaux, Université LavalAustralian Institute for Bioengineering and Nanotechnology, The University of Queensland
  • , Dominique MichaudAffiliated withCentre de Recherche et d’Innovation sur les Végétaux, Université Laval Email author 

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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.

Keywords

Clinically useful recombinant proteins Heterologous protein expression Recombinant protein degradation Companion protease inhibitors Protein stabilization