Heterologous expression of human H1 histones in yeast

被引:22
|
作者
Albig, W [1 ]
Runge, DM [1 ]
Kratzmeier, M [1 ]
Doenecke, D [1 ]
机构
[1] Univ Gottingen, Inst Biochem & Mol Zellbiol, D-37073 Gottingen, Germany
关键词
H1; histone; heterologous expression; Saccharomyces cerevisiae; two-dimensional gel electrophoresis; capillary zone electrophoresis;
D O I
10.1016/S0014-5793(98)01084-9
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:245 / 250
页数:6
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