Using single-molecule approach to visualize the nucleosome assembly in yeast nucleoplasmic extracts

• 来源期刊: 科学通报（英文版）
• 年，卷（期）: 2017,（6）
• 页码范围: 399-404
• 页数: 6页
• 语种: 英文

摘要

In eukaryotic cells,the smallest subunit of chromatin is the nucleosome,which consists of a segment of DNA wound on histone protein cores.Despite many years of effort,the process of nucleosome assembly and disassembly is still not very clear.Here,we present a convenient method to investigate the process of nucleosome assembly at the single molecule level.We invented a novel system derived from the yeast nucleoplasmic extracts (YNPE),and demonstrated that the YNPE supports the nucleosome assembly under physiological condition.By combining the total internal reflection fluorescence microscopy with microfluidic flow-cell technique,the dynamic process of nucleosome assembly in YNPE was visualized at single-molecule level.Our system provides a novel in vitro single-molecule tool to investigate the dynamics of nucleosome assembly under physiological conditions.