Chromatin Immunoprecipitation (ChiP) Protocol for the Analysis of Gene Regulation by Histone Modifications in Agave angustifolia Haw

Methods Mol Biol. 2018:1815:371-383. doi: 10.1007/978-1-4939-8594-4_26.

Abstract

Chromatin is a dynamic entity that regulates different biological processes crucial for the proper functioning of the cell. Chromatin regulation depends largely on the interactions that occur between DNA with histones and nonhistone proteins. The chromatin immunoprecipitation assay (ChiP) is a widely used technique for the study of these DNA-histone and DNA-nonhistone interactions and their biological repercussions. Here we describe a ChiP protocol that allows in vivo analysis of the associations of histone modifications with genomic DNA in Agave angustifolia Haw. Although this protocol is established for A. angustifolia, it can be used in other species to obtain similar results. We also propose a strategy to shorten the times in some steps of the standard protocol.

Keywords: Agave angustifolia; ChiP; H3K27me3; H3K4me3; LCYβ; PEPCase; RubS.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agave / genetics*
  • Agave / metabolism*
  • Chromatin / metabolism
  • Chromatin Immunoprecipitation / methods*
  • Cross-Linking Reagents / chemistry
  • Gene Expression Regulation, Plant*
  • Histones / metabolism*
  • Plant Leaves / metabolism
  • Polymerase Chain Reaction
  • Protein Processing, Post-Translational*
  • Solutions

Substances

  • Chromatin
  • Cross-Linking Reagents
  • Histones
  • Solutions