Abstract
A powerful method to identify binding sites in target genes is chromatin immunoprecipitation (ChIP), which allows the purification of in vivo formed complexes of a DNA-binding protein and associated DNA. Briefly, the method involves the fixation of plant tissue and the isolation of the total protein-DNA mixture, followed by an immunoprecipitation step with an antibody directed against the protein of interest and, subsequently, the DNA can be purified. Finally, the DNA can be analyzed by PCR for the enrichment of specific regions.
A drawback of ChIP is that for each protein another antibody is needed. To overcome this, a generic strategy is possible using tags fused to the protein of interest. In this case, only antibody is needed against the tag. This protocol describes the tagging of proteins and how to perform ChIP.
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Acknowledgments
This work was previously financed by the Netherlands Proteomics Centre (NPC) and now the work in de Folter laboratory is financed by the Mexican Science Council (CONACyT 82826) and CONCyTEG (08-03-K662-116).
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de Folter, S. (2011). Protein Tagging for Chromatin Immunoprecipitation from Arabidopsis . In: Pereira, A. (eds) Plant Reverse Genetics. Methods in Molecular Biology, vol 678. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-682-5_15
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DOI: https://doi.org/10.1007/978-1-60761-682-5_15
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