Jeronimo Célia, Poitras Christian, Robert François
Institut de recherches cliniques de Montréal, Montréal, Québec, Canada.
Département de médecine, Faculté de médecine, Université de Montréal, Montréal, Québec, Canada.
Methods Mol Biol. 2025;2919:155-177. doi: 10.1007/978-1-0716-4486-7_9.
Mapping the position of nucleosomes in vivo is key to our understanding of chromatin-based processes such as gene transcription, DNA replication, and repair. Methods based on micrococcal nuclease (MNase) digestion of chromatin are widely used to map nucleosomes, but these methods suffer from some limitations. More recently, nucleosome mapping methods that take advantage of our ability to convert cysteine residues-carefully inserted in histone proteins-into nucleases provide alternatives to MNase-based assays. Here, we provide a detailed protocol for the mapping of nucleosomes via H3Q85C-directed DNA cleavage in Saccharomyces cerevisiae.
绘制体内核小体的位置对于我们理解基于染色质的过程（如基因转录、DNA复制和修复）至关重要。基于微球菌核酸酶（MNase）消化染色质的方法被广泛用于绘制核小体，但这些方法存在一些局限性。最近，利用我们将精心插入组蛋白中的半胱氨酸残基转化为核酸酶的能力的核小体映射方法为基于MNase的检测提供了替代方案。在这里，我们提供了一种在酿酒酵母中通过H3Q85C定向DNA切割绘制核小体的详细方案。
