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Preparation of Chromatin Assembly Extracts from Preblastoderm Drosophila Embryos

  • Edgar Bonte
  • Peter B. Becker
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 523)

Abstract

A rigorous biochemical analysis of chromatin structure and function requires the assembly of chromatin in vitro. A useful alternative to reconstituting nucleosomal arrays from pure or recombinant histones by salt gradient dialysis is the assembly of more complex chromatin from assembly extracts under physiological conditions. Extracts from preblastoderm embryos have proven to be particularly efficient, due to the presence of large stores of native complexes of histones, histone chaperones and ATP-dependent nucleosome spacing factors. The resulting chromatin is an excellent approximation of physiological chromatin in vivo. This chapter describes the preparation of chromatin assembly extracts and the chromatin assembly reaction.

Key words

Nucleosome assembly chromatin reconstitution in vitro analysis linker histone Drosophila 

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Edgar Bonte
    • 1
  • Peter B. Becker
    • 1
  1. 1.MolekularbiologieAdolf-Butenandt-InstitutMünchenGermany

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