Heterologous expression of human H1 histones in yeast

FEBS Letters

Albig, Werner¹  Runge, Dorothee Monika¹  Doenecke, Detlef¹  Kratzmeier, Martin¹ 

[1]Institut für Biochemie und Molekulare Zellbiologie, Georg-August-Universität Göttingen, Humboldtallee 23, D-37073 Göttingen, Germany

关键词: H1 histone;    Heterologous expression;    Saccharomyces cerevisiae;    Two-dimensional gel electrophoresis;    Capillary zone electrophoresis;    APS;    ammonium persulfate;    PCA;    perchloric acid;    TCA;    trichloroacetic acid;    CZE;    capillary zone electrophoresis;    RPMI;    Roswell Park Memorial Institute medium;    HPMC;    hydroxypropyl-methylcellulose;    PBS;    phosphate buffered saline;    SDS-PAGE;    SDS-polyacrylamide gel electrophoresis;    AUT gel;    acid-urea-triton electrophoresis gel;    TEMED;    N;    N;    N′;    N′-tetramethylethylenediamine;   

DOI  :  10.1016/S0014-5793(98)01084-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The complete set of seven human H1 histone subtype genes was heterologously expressed in yeast. Since Saccharomyces cerevisiae lacks standard histone H1 we 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 was barely possible until now.

【 授权许可】

Unknown