title: Studying Transcription Factor Binding to Specific Genomic Loci by Chromatin Immunoprecipitation (ChIP).
abstract: Plant hormone signaling involves complex transcriptional networks, where transcription factors orchestrate the control of specific gene expression. These networks include cross talk between hormone signaling pathways, and the integration of environmental signals and the developmental program. Understanding how particular transcription factors respond and integrate specific signals is crucial in order to understand the basic mechanisms of hormonal signaling and cross talk. Studying transcription factor binding at specific genomic loci by chromatin immunoprecipitation (ChIP) is therefore a valuable technique in order to analyze transcriptional regulation. The method is based on cross-linking proteins to DNA, the isolation of chromatin, and immunoprecipitation of a transcription factor of interest. The attached DNA is then recovered and analyzed by quantitative real-time PCR in order to establish binding sites of the respective transcription factor. Here, we present a relatively simple and short protocol for ChIP on single loci.
authors.affiliation: Cell and Developmental Biology, John Innes Centre, Norwich, NR4 7UH, UK.
authors.forename: S Vinod
authors.initials: SV
authors.lastname: Kumar
authors.affiliation: Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences (BOKU), Muthgasse 18, A-1190, Vienna, Austria. doris.lucyshyn@boku.ac.at.
authors.forename: Doris
authors.initials: D
authors.lastname: Lucyshyn
mesh_terms: D001665:Binding Sites / Q000502:physiology; D002843:Chromatin / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation / Q000379:methods; D004247:DNA / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology; D023281:Genomics / Q000379:methods; D010937:Plant Growth Regulators / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology*; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D002843:Chromatin; D010937:Plant Growth Regulators; D010940:Plant Proteins; D014157:Transcription Factors; D004247:DNA
keywords: Binding site; Chromatin extraction; Chromatin immunoprecipitation (ChIP); Gene expression; Transcription factor