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Cytoskeleton Methods and Protocols pp 207–225Cite as

Purification and Localization of Intraflagellar Transport Particles and Polypeptides

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 586))

Summary

The growth and maintenance of almost all cilia and flagella are dependent on the proper functioning of the process of intraflagellar transport (IFT). This includes the primary cilia of most cells in humans that are in interphase or the Go phase of the cell cycle. The model system for the study of IFT is the flagella of the bi-flagellate green alga Chlamydomonas. It is in this organism that IFT was first discovered, and genetic data from a Chlamydomonas mutant first linked the process of IFT to polycystic kidney disease in humans. The information given in this chapter addresses procedures to purify IFT particles from flagella and localize these particles, and their associated motor proteins, in flagella using light and electron microscopic approaches.

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Notes

  1. 1.

    The term ‘flagella’ will be used throughout this manuscript, but readers should realize that the process of IFT occurs—and is just as important—in cilia as well. Indeed, many developmental problems and diseases of humans are related to defects in primary cilia, some of which derive from defects in IFT (see reference ( 1 ) R.D. Sloboda, and J.L. Rosenbaum, Making sense of cilia and flagella. J Cell Biol 179 (2007) 575–82. for a recent overview of this field).

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Acknowledgments

Work in the author’s lab is supported by the NSF (MCB 0418877) and the NIH (DK 071720). This support is greatly appreciated.

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Authors and Affiliations

  1. Department of Biological Sciences, Dartmouth College, Hanover, NH, USA

    Roger D. Sloboda

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  1. Roger D. Sloboda
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Editors and Affiliations

  1. Brooklyn College, City University of New York, Bedford Ave. 2900, New York, 11210-2889, U.S.A.

    Ray H. Gavin

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Sloboda, R.D. (2009). Purification and Localization of Intraflagellar Transport Particles and Polypeptides. In: Gavin, R. (eds) Cytoskeleton Methods and Protocols. Methods in Molecular Biology, vol 586. Humana Press. https://doi.org/10.1007/978-1-60761-376-3_11

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  • DOI: https://doi.org/10.1007/978-1-60761-376-3_11

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-375-6

  • Online ISBN: 978-1-60761-376-3

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