Ferraro Aileen R, Lewis Zachary A
Department of Microbiology, University of Georgia, Athens, GA, USA.
Methods Mol Biol. 2018;1775:241-250. doi: 10.1007/978-1-4939-7804-5_19.
Chromatin immunoprecipitation paired with next-generation sequencing (ChIP-seq) can be used to determine genome-wide distribution of transcriptions factors, transcriptional machinery, or histone modifications. DNA-protein interactions are covalently cross-linked with the addition of formaldehyde. Chromatin is prepared and sheared, then immunoprecipitated with the appropriate antibody. After reversal of cross-linking and treating with protease, the resulting DNA fragments are sequenced and mapped to the reference genome to determine overall enrichment. Here we describe a method of ChIP-seq for investigating protein-DNA interactions in the filamentous fungus Neurospora crassa.
染色质免疫沉淀与下一代测序技术相结合（ChIP-seq）可用于确定转录因子、转录机制或组蛋白修饰在全基因组范围内的分布。通过添加甲醛使DNA与蛋白质的相互作用发生共价交联。制备并剪切染色质，然后用适当的抗体进行免疫沉淀。在交联逆转并经蛋白酶处理后，对所得的DNA片段进行测序，并将其定位到参考基因组以确定总体富集情况。在此，我们描述了一种用于研究丝状真菌粗糙脉孢菌中蛋白质-DNA相互作用的ChIP-seq方法。
