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Knocking Out the Wall: Protocols for Gene Targeting in Physcomitrella patens

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The Plant Cell Wall

Part of the book series: Methods in Molecular Biology ((MIMB,volume 715))

Abstract

The moss Physcomitrella patens has become established as a model for investigating plant gene function due to the feasibility of gene targeting. The chemical composition of the P. patens cell wall is similar to that of vascular plants and phylogenetic analyses of glycosyltransferase sequences from the P. patens genome have identified genes that putatively encode cell wall biosynthetic enzymes, providing a basis for investigating the evolution of cell wall polysaccharides and the enzymes that synthesize them. The protocols described in this chapter provide methods for targeted gene knockout in P. patens, from constructing vectors and maintaining cultures to transforming protoplasts and analyzing the genotypes and phenotypes of the resulting transformed lines.

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Acknowledgements

We thank the members of the moss community for their collegiality and helpful suggestions. Magdalena Bezanilla and the members of her group provided invaluable advice and assistance. We thank Didier Schaefer for the gift of pBHSNR. This project was supported by the National Research Initiative Competitive Grant no. 2007-35318-18389 from the USDA National Institute of Food and Agriculture.

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Authors and Affiliations

  1. Department of Biological Sciences, Center for Biotechnology and Life Sciences, University of Rhode Island, Kingston, RI, USA

    Alison W. Roberts, Christos S. Dimos, Michael J. Budziszek Jr., Chessa A. Goss & Virginia Lai

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  1. Alison W. Roberts
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  2. Christos S. Dimos
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  3. Michael J. Budziszek Jr.
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  4. Chessa A. Goss
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  5. Virginia Lai
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Editors and Affiliations

  1. School of Natural Sciences, Dept. of Botany & Plant Science, National University of Ireland, Galway, University Road, Galway, Ireland

    Zoë A. Popper

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Roberts, A.W., Dimos, C.S., Budziszek, M.J., Goss, C.A., Lai, V. (2011). Knocking Out the Wall: Protocols for Gene Targeting in Physcomitrella patens . In: Popper, Z. (eds) The Plant Cell Wall. Methods in Molecular Biology, vol 715. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-008-9_19

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  • DOI: https://doi.org/10.1007/978-1-61779-008-9_19

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-007-2

  • Online ISBN: 978-1-61779-008-9

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