A Yeast Chromatin-enriched Fractions Purification Approach, yChEFs, from Saccharomyces cerevisiae

被引:3
|
作者
Cuevas-Bermudez, Abel [1 ]
Garrido-Godino, Ana, I [1 ]
Gutierrez-Santiago, Francisco [1 ]
Navarro, Francisco [1 ,2 ]
机构
[1] Univ Jaen, Dept Biol Expt Genet, Jaen, Spain
[2] Univ Jaen, Ctr Estudios Avanzados Aceite Oliva & Olivar, Paraje Las Lagunillas S-N, Jaen 23071, Spain
来源
BIO-PROTOCOL | 2020年 / 10卷 / 01期
关键词
Chromatin; Proteins; Yeast; Mass spectrometry; Saccharomyces cerevisiae; DNA-REPLICATION; CELL-CYCLE; PROTEINS; PROTEOME;
D O I
10.21769/BioProtoc.3471
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
We have adapted a previous procedure and improved an approach that we named yChEFs (yeast Chromatin Enriched Fractions) for purifying chromatin fractions. This methodology allows the easy, reproducible and scalable recovery of proteins associated with chromatin. By using yChEFs, we bypass subcellular fractionation requirements involved when using zymolyase to obtain the spheroplast, which is employed in many other procedures. Employing small amount of culture cells and small volumes of solutions during the yChEFs procedure is very useful to allow many samples to be handled at the same time, and also reduces costs and efforts. The purified proteins associated with chromatin fractions obtained by yChEFs can be analyzed by Western blot (Figure 1) or combined with mass spectrometry for proteomic analyses.
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页数:9
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