摘要
为了制备体外酵母DNA序列组装核小体所需的组蛋白,利用酸抽提方法从未经饥饿处理和经过不同时间饥饿处理的酿酒酵母细胞中分离组蛋白,经SDS-PAGE电泳分析和Bradford法测定蛋白浓度,发现抽提物中含有组蛋白H1、H2A、H2B、H3和H4,电泳条带位置正确、纯度较高,正常细胞的抽提物中蛋白总量达到158μg/mL。试验结果表明该方法可以提取出较高质量的酿酒酵母组蛋白。
The work has provided a procedure to obtain the nuclear proteins that are required for assembly of chromatin template in vitro.Histones were extracted from untreated and starving yeast cells by acid extraction method.The qualities of histones were analyzed by SDS-PAGE electrophoresis and Bradford method.Histones with satisfactory stability were isolated properly and the protein concentrations were 158 μg/mL.The results indicated that this method had the ability to extract histones with high purity and concentration from Saccharomyces cerevisiae.The work was the basic of studying chromatin assembly in vitro.
出处
《生物技术通报》
CAS
CSCD
北大核心
2011年第11期225-228,共4页
Biotechnology Bulletin
基金
国家自然科学基金资助项目(61072129)
内蒙古科技大学创新基金项目(2010NC056)
关键词
酿酒酵母
组蛋白提取
饥饿细胞
核小体定位
体外组装
Saccharomyces cerevisiaet Extraction of histones Starvation cell Nucleosome positioning Chromatin assembly
