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Chromatin Immunoprecipitation for Studying Transcriptional Regulation in Xenopus Oocytes and Tadpoles

  • David Stewart
  • Akihiro Tomita
  • Yun-Bo Shi
  • Jiemin Wong
Part of the Methods in Molecular Biology™ book series (MIMB, volume 322)

Abstract

Understanding the accurate temporal and spatial regulation of gene expression during development requires knowledge of the spectrum of transcription factors and cofactors involved and their functional interplay with chromatin. Chromatin immunoprecipitation (ChIP) has become a powerful technique that allows us to do so. A typical ChIP assay involves (1) treating cells or tissues with formaldehyde to rapidly crosslink chromatin-associated proteins to DNA, (2) shearing chromatin by sonication into small fragments, (3) immunoprecipitation of the proteins of interest, (4) reversal of crosslinking, and (5) quantitating the specific associated DNA sequences by PCR. Here we present and discuss the protocols we have developed over the years for ChIP assays using Xenopus oocytes and tadpole tissues as experimental materials.

Key Words

Chromatin immunoprecipitation (ChIP) tadpole thyroid hormone receptor transcription Xenopus oocyte 

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Copyright information

© Humana Press Inc., Totowa, NJ 2006

Authors and Affiliations

  • David Stewart
    • 1
  • Akihiro Tomita
    • 2
  • Yun-Bo Shi
    • 2
  • Jiemin Wong
    • 1
  1. 1.Department of Molecular and Cellular BiologyBaylor College of MedicineHouston
  2. 2.Section on Molecular Morphogenesis, Laboratory of Gene Regulation and DevelopmentNational Institute of Child Health and Human Disease, National Institutes of HealthBethesda

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