Abstract
Chromatin immunoprecipitation (ChIP) is used to map the interaction between proteins and DNA at a specific genomic locus in the living cell. The protein–DNA complexes are stabilized already in vivo by reversible crosslinking and the DNA is sheared by sonication or enzymatic digestion into fragments suitable for the subsequent immunoprecipitation step. Antibodies recognizing chromatin-linked proteins, transcription factors, artificial tags, or specific protein modifications are then used to pull down DNA–protein complexes containing the target. After reversal of crosslinks and DNA purification locus-specific quantitative PCR is used to determine the amount of DNA that was associated with the target at a given time point and experimental condition. DNA quantification can be carried out for several genomic regions by multiple qPCRs or at a genome-wide scale by massive parallel sequencing (ChIP-Seq).
These two authors contributed equally to the work
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Acknowledgments
ChIP development and optimization in A. nidulans was supported by grants from the Austrian Science Fund P19731-B11 and Vienna Science and Technology Fund—WWTF—Project LS 09-042. The adaptation to Fusarium species was carried out in the frame of the Austrian Science Fund “Special Research AreA” Project SFB–S10003.
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Boedi, S., Reyes-Dominguez, Y., Strauss, J. (2012). Chromatin Immunoprecipitation Analysis in Filamentous Fungi. In: Keller, N., Turner, G. (eds) Fungal Secondary Metabolism. Methods in Molecular Biology, vol 944. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-122-6_16
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DOI: https://doi.org/10.1007/978-1-62703-122-6_16
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