Cell cycle synchronization in xenopus egg extracts

Peter J. Gillespie, Julia Neusiedler, Kevin Creavin, Gaganmeet Singh Chadha, John Blow (Lead / Corresponding author)

    Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)peer-review

    6 Citations (Scopus)

    Abstract

    Many important discoveries in cell cycle research have been made using cell-free extracts prepared from the eggs of the South African clawed frog Xenopus laevis . These extracts effi ciently support the key nuclear functions of the eukaryotic cell cycle in vitro under apparently the same controls that exist in vivo. The Xenopus cell-free system is therefore uniquely suited to the study of the mechanisms, dynamics and integration of cell cycle regulated processes at a biochemical level. Here, we describe methods currently in use in our laboratory for the preparation of Xenopus egg extracts and demembranated sperm nuclei. We detail how these extracts can be used to study the key transitions of the eukaryotic cell cycle and describe conditions under which these transitions can be manipulated by addition of drugs that either retard or advance passage. In addition, we describe in detail essential techniques that provide a practical starting point for investigating the function of proteins involved in the operation of the eukaryotic cell cycle.

    Original languageEnglish
    Title of host publicationCell cycle oscillators
    Subtitle of host publicationmethods and protocols
    EditorsAmanda S. Coutts, Louise Weston
    Place of PublicationNew York
    PublisherHumana Press
    Pages101-147
    Number of pages47
    ISBN (Electronic)9781493929573
    ISBN (Print)9781493929566
    DOIs
    Publication statusPublished - 2016

    Publication series

    NameMethods in Molecular Biology
    PublisherSpringer New York
    Volume1342
    ISSN (Print)1064-3745

    Keywords

    • Cell cycle
    • Cell-free system
    • DNA replication
    • Egg extract
    • In vitro
    • Synchronization
    • Xenopus

    ASJC Scopus subject areas

    • Molecular Biology
    • Genetics

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