Protocol: Chromatin immunoprecipitation (ChIP) methodology to investigate histone modifications in two model diatom species. - Archive ouverte HAL Access content directly
Journal Articles Plant Methods Year : 2012

Protocol: Chromatin immunoprecipitation (ChIP) methodology to investigate histone modifications in two model diatom species.

(1) , (1) , (1)
1

Abstract

ABSTRACT: In this report we describe a chromatin immunoprecipitation (ChIP) protocol for two fully sequenced model diatom species Phaeodactylum tricornutum and Thalassiosira pseudonana. This protocol allows the extraction of satisfactory amounts of chromatin and gives reproducible results. We coupled the ChIP assay with real time quantitative PCR. Our results reveal that the two major histone marks H3K4me2 and H3K9me2 exist in P. tricornutum and T. pseudonana. As in other eukaryotes, H3K4me2 marks active genes whereas H3K9me2 marks transcriptionally inactive transposable elements. Unexpectedly however, T. pseudonana housekeeping genes also show a relative enrichment of H3K9me2. We also discuss optimization of the procedure, including growth conditions, cross linking and sonication. Validation of the protocol provides a set of genes and transposable elements that can be used as controls for studies using ChIP in each diatom species. This protocol can be easily adapted to other diatoms and eukaryotic phytoplankton species for genetic and biochemical studies.
Fichier principal
Vignette du fichier
1746-4811-8-48.pdf (475.5 Ko) Télécharger le fichier
Vignette du fichier
1746-4811-8-48.xml (62.17 Ko) Télécharger le fichier
Origin : Publisher files allowed on an open archive
Format : Other
Loading...

Dates and versions

inserm-00776647 , version 1 (15-01-2013)

Identifiers

Cite

Xin Lin, Leïla Tirichine, Chris Bowler. Protocol: Chromatin immunoprecipitation (ChIP) methodology to investigate histone modifications in two model diatom species.. Plant Methods, 2012, 8 (1), pp.48. ⟨10.1186/1746-4811-8-48⟩. ⟨inserm-00776647⟩
142 View
291 Download

Altmetric

Share

Gmail Facebook Twitter LinkedIn More