Protocol: Chromatin immunoprecipitation (ChIP) methodology to investigate histone modifications in two model diatom species. - Inserm - Institut national de la santé et de la recherche médicale Accéder directement au contenu
Article Dans Une Revue Plant Methods Année : 2012

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

Résumé

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
1746-4811-8-48.xml (62.17 Ko) Télécharger le fichier
Origine : Fichiers éditeurs autorisés sur une archive ouverte
Format : Autre
Loading...

Dates et versions

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

Identifiants

Citer

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⟩
149 Consultations
303 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More