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Isolation and Characterization of Ice-Binding Proteins from Higher Plants

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Plant Cold Acclimation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1166))

Abstract

The characterization of ice-binding proteins from plants can involve many techniques, only a few of which are presented here. Chief among these methods are tests for ice recrystallization inhibition activity. Two distinct procedures are described; neither is normally used for precise quantitative assays. Thermal hysteresis assays are used for quantitative studies but are also useful for ice crystal morphologies, which are important for the understanding of ice-plane binding. Once the sequence of interest is cloned, recombinant expression, necessary to verify ice-binding protein identity can present challenges, and a strategy for recovery of soluble, active protein is described. Lastly, verification of function in planta borrows from standard protocols, but with an additional screen applicable to ice-binding proteins. Here we have attempted to assist researchers wishing to isolate and characterize ice-binding proteins from plants with a few methods critical to success.

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Acknowledgements

We would like to thank Dr. M. Kuiper, along with many undergraduate students who have participated in data collection and “trouble-shooting” these techniques over the years. The research was supported by a CIHR and NSERC (Canada) grants to PLD and VKW, respectively.

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Authors and Affiliations

  1. Department of Biomedical and Molecular Sciences, Queen’s University, Kingston, ON, Canada, K7L 3N6

    Adam J. Middleton, Peter L. Davies & Virginia K. Walker

  2. Department of Biochemistry, University of Otago, Dunedin, 9016, New Zealand

    Adam J. Middleton

  3. Department of Biology, Queen’s University, Kingston, ON, Canada, K7L 3N6

    Barbara Vanderbeld, Melissa Bredow, Heather Tomalty, Peter L. Davies & Virginia K. Walker

Authors
  1. Adam J. Middleton
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  2. Barbara Vanderbeld
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  3. Melissa Bredow
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  4. Heather Tomalty
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  5. Peter L. Davies
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  6. Virginia K. Walker
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Corresponding author

Correspondence to Virginia K. Walker .

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Editors and Affiliations

  1. Max-Planck-Institut für Molekulare Pflanzenphysiologie, Potsdam, Germany

    Dirk K. Hincha

  2. Max-Planck-Institut für Molekulare Pflanzenphysiologie, Potsdam, Germany

    Ellen Zuther

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Middleton, A.J., Vanderbeld, B., Bredow, M., Tomalty, H., Davies, P.L., Walker, V.K. (2014). Isolation and Characterization of Ice-Binding Proteins from Higher Plants. In: Hincha, D., Zuther, E. (eds) Plant Cold Acclimation. Methods in Molecular Biology, vol 1166. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0844-8_19

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  • DOI: https://doi.org/10.1007/978-1-4939-0844-8_19

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0843-1

  • Online ISBN: 978-1-4939-0844-8

  • eBook Packages: Springer Protocols

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