Expression and purification of human heat-shock transcription factor 1

被引：24

作者：

Soncin, F ^{[1
]}

Prevelige, R ^{[1
]}

Calderwood, SK ^{[1
]}

机构：

[1] HARVARD UNIV,SCH MED,DANA FARBER CANC INST,JOINT CTR RADIAT THERAPY,BOSTON,MA 02115

来源：

PROTEIN EXPRESSION AND PURIFICATION | 1997年 / 9卷 / 01期

关键词：

D O I：

10.1006/prep.1996.0672

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Heat-shock factor 1 (HSF1) is a 57-kDa cytoplasmic protein which binds to the promoters of heat-shock genes and activates transcription during heat shock. We describe here the expression and purification of the 529 amino acids form of human HSF1. We designed a new and complete purification protocol involving ammonium sulfate precipitation, heparin-Sepharose affinity, and ion-exchange chromatography, which allows the purification of large amounts of pure and active recombinant protein. HSF1 isolated by this method is pure as that assessed by SDS-PAGE and reverse-phase HPLC. The purified protein is recognized by specific anti-HSF1 antibodies, binds to heat-shock elements, and activates the promoter of the heat-shock protein 70A gene in vitro. (C) 1997 Academic Press.

引用

页码：27 / 32

页数：6

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