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Xenopus Protocols pp 301–314Cite as

Use of Intact Xenopus Oocytes in Nucleocytoplasmic Transport Studies

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 322))

Abstract

Because of its large nucleus, the Xenopus laevis oocyte offers an excellent system to study nucleocytoplasmic transport. This system, in combination with electron microscopy, has provided much of our insight into the mechanisms of nuclear import and export. In a typical experiment, the nuclear transport substrate is first labeled with colloidal gold, and the resulting complex is injected into the cytoplasm (to study nuclear import) or the nucleus (to study nuclear export) of Xenopus oocytes. The oocytes are then fixed, dehydrated, infiltrated, and embedded into an epoxy resin. Following resin polymerization, thin sections of oocyte nuclei are obtained and examined under an electron microscope. Subsequent evaluation of the position and distribution of the gold-labeled substrate reveals whether the substrate has undergone nuclear import (or export) and the position of rate-limiting events. This chapter describes in detail the protocols for performing electron microscopy import assays with Xenopus oocytes and presents some data illustrating the types of experiments possible using this system.

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Author information

Authors and Affiliations

  1. Department of Zoology, University of British Columbia, Vancouver, Columbia, Canada

    Nelly Panté

Authors
  1. Nelly Panté
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Editor information

Editors and Affiliations

  1. Ottawa Health Research Institute, Ottawa Hospital, Ottawa, Ontario, Canada

    X. Johné Liu

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© 2006 Humana Press Inc., Totowa, NJ

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Panté, N. (2006). Use of Intact Xenopus Oocytes in Nucleocytoplasmic Transport Studies. In: Liu, X.J. (eds) Xenopus Protocols. Methods in Molecular Biology™, vol 322. Humana Press. https://doi.org/10.1007/978-1-59745-000-3_21

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  • DOI: https://doi.org/10.1007/978-1-59745-000-3_21

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-362-6

  • Online ISBN: 978-1-59745-000-3

  • eBook Packages: Springer Protocols

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