Galdieri Luciano, Moon John, Vancura Ales
Department of Biological Sciences, St. John's University, Queens, NY, USA.
Methods Mol Biol. 2012;809:255-65. doi: 10.1007/978-1-61779-376-9_17.
Histone acetylation is the most studied posttranslation modification of nucleosomes. Understanding the mechanisms involved in global and promoter-specific histone acetylation will shed light on the control of transcriptional regulation. Chromatin immunoprecipitation is a powerful technique to study protein-DNA interactions in vivo. Proteins and DNA are cross-linked with formaldehyde, cells are lysed, and DNA is sheared by sonication. Protein-DNA complexes are immunoprecipitated with antibodies specific for total and acetylated histones and the relative occupancy of acetylated and total histones at selected loci is assessed by real-time PCR of the purified DNA.
组蛋白乙酰化是对核小体研究最多的翻译后修饰。了解全局和启动子特异性组蛋白乙酰化所涉及的机制将有助于揭示转录调控的控制。染色质免疫沉淀是一种在体内研究蛋白质 - DNA相互作用的强大技术。蛋白质和DNA用甲醛交联，细胞裂解，DNA通过超声破碎。蛋白质 - DNA复合物用针对总组蛋白和乙酰化组蛋白的特异性抗体进行免疫沉淀，通过对纯化DNA的实时PCR评估选定基因座处乙酰化组蛋白和总组蛋白的相对占有率。
