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PCR-Based Bioprospecting for Homing Endonucleases in Fungal Mitochondrial rRNA Genes

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Homing Endonucleases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1123))

Abstract

Fungal mitochondrial genomes act as “reservoirs” for homing endonucleases. These enzymes with their DNA site-specific cleavage activities are attractive tools for genome editing and gene therapy applications. Bioprospecting and characterization of naturally occurring homing endonucleases offers an alternative to synthesizing artificial endonucleases. Here, we describe methods for PCR-based screening of fungal mitochondrial rRNA genes for homing endonuclease encoding sequences, and we also provide protocols for the purification and biochemical characterization of putative native homing endonucleases.

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Acknowledgments

This work is supported by a Discovery grant from the Natural Sciences and Engineering Research Council of Canada (NSERC) to G.H. M.H. is supported by the Egyptian Ministry of Higher Education and Scientific Research; and T.K.G. and C.S. would like to acknowledge funding support from the Faculty of Science Graduate Award program (University of Manitoba).

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Authors and Affiliations

  1. Department of Biochemistry, Université de Montréal, Montréal, QC, Canada

    Mohamed Hafez

  2. Department of Microbiology, University of Manitoba, Winnipeg, MB, Canada

    Tuhin Kumar Guha, Chen Shen, Jyothi Sethuraman & Georg Hausner

Authors
  1. Mohamed Hafez
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  2. Tuhin Kumar Guha
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  3. Chen Shen
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  4. Jyothi Sethuraman
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  5. Georg Hausner
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Editors and Affiliations

  1. University of Western Ontario, London, Ontario, Canada

    David R. Edgell

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Hafez, M., Guha, T.K., Shen, C., Sethuraman, J., Hausner, G. (2014). PCR-Based Bioprospecting for Homing Endonucleases in Fungal Mitochondrial rRNA Genes. In: Edgell, D. (eds) Homing Endonucleases. Methods in Molecular Biology, vol 1123. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-968-0_3

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  • DOI: https://doi.org/10.1007/978-1-62703-968-0_3

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-967-3

  • Online ISBN: 978-1-62703-968-0

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