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)

5 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

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  • Cite this

    Gillespie, P. J., Neusiedler, J., Creavin, K., Chadha, G. S., & Blow, J. (2016). Cell cycle synchronization in xenopus egg extracts. In A. S. Coutts, & L. Weston (Eds.), Cell cycle oscillators: methods and protocols (pp. 101-147). (Methods in Molecular Biology; Vol. 1342). Humana Press. https://doi.org/10.1007/978-1-4939-2957-3_6