Unnikrishnan Ashwin, Akiyoshi Bungo, Biggins Sue, Tsukiyama Toshio
Basic Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
Methods Mol Biol. 2012;833:115-23. doi: 10.1007/978-1-61779-477-3_8.
We have recently established a system for purifying minichromosomes in a native state from Saccharomyces cerevisiae. This system is extremely efficient, and a single-step purification yields samples with sufficient purity and quantity for mass spectrometry (MS) analysis of histones and non-histone proteins tightly associated with the minichromosome. The templates can also be used in various biochemical assays in vitro, such as transcription and recombination, and could be suitable for EM or other biophysical studies.
我们最近建立了一个从酿酒酵母中以天然状态纯化微型染色体的系统。该系统效率极高,单步纯化就能得到纯度和数量足以用于对与微型染色体紧密相关的组蛋白和非组蛋白进行质谱(MS)分析的样品。这些模板还可用于各种体外生化分析,如转录和重组分析,并且可能适用于电子显微镜(EM)或其他生物物理研究。