Yeast Nucleoplasmic Extracts and an Application to Visualize Chromatin Assembly on Single Molecules of DNA.
Yeast Nucleoplasmic Extracts and an Application to Visualize Chromatin Assembly on Single Molecules of DNA.
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DOI：
10.1007/978-1-0716-0868-5_15
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发表时间：
2021
影响因子：
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通讯作者：
Yong Wang;Y. Fu
中科院分区：
文献类型：
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作者：
Yong Wang;Y. Fu
在真核细胞中，基因组DNA被包装到染色质中，染色质的基本单位是核小体。由于研究方法的局限性，在生理条件下研究染色质形成的机制具有内在的困难。在这里，我们描述了如何准备一个生化系统称为酵母核质提取物（YNPE）。YNPE来源于酵母细胞核，体外系统可以模拟体内酵母细胞核的生理条件。在YNPE中，利用全内反射荧光显微镜在单分子水平上真实的实时观察了染色质组装的动态过程。YNPE为研究生理条件下染色质组装的许多方面提供了一种新的工具，并适用于单分子方法。
In eukaryotic cells, the genomic DNA is packaged into chromatin, the basic unit of which is the nucleosome. Studying the mechanism of chromatin formation under physiological conditions is inherently difficult due to the limitations of research approaches. Here we describe how to prepare a biochemical system called yeast nucleoplasmic extracts (YNPE). YNPE is derived from yeast nuclei, and the in vitro system can mimic the physiological conditions of the yeast nucleus in vivo. In YNPE, the dynamic process of chromatin assembly has been observed in real time at the single-molecule level by total internal reflection fluorescence microscopy. YNPE provides a novel tool to investigate many aspects of chromatin assembly under physiological conditions and is competent for single-molecule approaches.