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Generating Transgenic Frog Embryos by Restriction Enzyme Mediated Integration (REMI)

  • Shoko Ishibashi
  • Kristen L. KrollEmail author
  • Enrique AmayaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 917)

Abstract

Here we present a protocol for generating transgenic embryos in Xenopus laevis and Xenopus tropicalis. The method includes three steps: (1) The preparation of high-speed egg extracts, which facilitates the replacement of protamines in sperm nuclei with nucleosomes and decondenses the chromatin of sperm nuclei; (2) The isolation of sperm nuclei; and (3) The mixing of sperm nuclei, restriction enzyme, and high-speed extract in vitro, following by nuclear transplantation into unfertilized eggs to generate the transgenic embryos. This procedure generates non-mosaic transgenic embryos at high frequency and efficiency.

Key words

Xenopus Restriction enzyme mediated integration Transgenesis 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.The Healing Foundation Centre, The Faculty of Life SciencesUniversity of ManchesterManchesterEngland, UK
  2. 2.Department of Developmental BiologyWashington University School of MedicineSt LouisUSA
  3. 3.Healing Foundation Centre, Faculty of Life SciencesUniversity of ManchesterManchesterEngland, UK

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