Heterologous expression of human H1 histones in yeast
Heterologous expression of human H1 histones in yeast
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DOI：
10.1016/s0014-5793(98)01084-9
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发表时间：
1998-09-18
期刊：
影响因子：
3.5
通讯作者：
Doenecke, D
中科院分区：
文献类型：
--
作者：
Albig, W;Runge, DM;Doenecke, D
人H1组蛋白7个完整亚型基因在酵母中异源表达。由于酿酒酵母缺乏标准的组蛋白H1，我们可以在不受内源性H1组蛋白污染的情况下，以纯形式分离出重组表达的人H1亚型，该表达系统中H1组蛋白的分离不需要标记，用5% （0.74 M）高氯酸一次提取就可以提取出具有真实初级结构的亚型。分离得到的H1组蛋白分别经过二维凝胶电泳和毛细管区带电泳，将亚型基因定位到相应的蛋白点或蛋白峰上。这使我们能够将转录数据与蛋白质数据联系起来，这在1998年欧洲生化学会联合会之前几乎是不可能的。
The complete set of seven human H1 histone subtype genes was heterologously expressed in yeast. Since Saccharomyces cerevisiae lacks standard histone H1 me could isolate each recombinantly expressed human H1 subtype in pure form without contamination by endogenous H1 histones, For isolation of the H1 histones in this expression system no tagging was needed and the isoforms could be extracted with the authentic primary structure by a single extraction step with 5% (0.74 M) perchloric acid. The isolated H1 histone proteins were used to assign the subtype genes to the corresponding protein spots or peaks after two-dimensional gel electrophoresis and capillary zone electrophoresis, respectively. This allowed us to correlate transcriptional data with protein data, which mas barely possible until nom, (C) 1998 Federation of European Biochemical Societies.