Chapter title |
μChIP-Seq for Genome-Wide Mapping of In Vivo TF-DNA Interactions in Arabidopsis Root Protoplasts
|
---|---|
Chapter number | 19 |
Book title |
Root Development
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7747-5_19 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7746-8, 978-1-4939-7747-5
|
Authors |
Para, Alessia, Li, Ying, Coruzzi, Gloria M, Alessia Para, Ying Li, Gloria M. Coruzzi |
Abstract |
Chromatin immunoprecipitation (ChIP) is a widely used method to map the position of DNA-binding proteins such as histones and transcription factors (TFs) upon their interaction with particular regions of the genome. To examine the genomic distribution of a TF in specific cell types in response to a change in nitrogen concentration, we developed a micro-ChIP (μChIP) protocol that requires only ~5000 Arabidopsis cells transiently expressing the Arabidopsis TF Basic Leucine Zipper 1 (bZIP1) fused to the glucocorticoid receptor (GR) domain that mediates nuclear import in the presence of dexamethasone. The DNA fragments obtained from the immunoprecipitation of bZIP1-DNA complexes were analyzed by next-generation sequencing (ChIP-seq), which helped uncover genome-wide associations between a bZIP1 and its targets in plant cells upon fluctuations in nitrogen availability. |
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