A RAPID ONE-STEP METHOD TO PURIFY BACULOVIRUS-EXPRESSED HUMAN ESTROGEN-RECEPTOR TO BE USED IN THE ANALYSIS OF THE OXYTOCIN PROMOTER

被引：14

作者：

BEEKMAN, JM ^{[1
]}

COONEY, AJ ^{[1
]}

ELLISTON, JF ^{[1
]}

TSAI, SY ^{[1
]}

TSAI, MJ ^{[1
]}

机构：

[1] BAYLOR COLL MED,DEPT CELL BIOL,HOUSTON,TX 77030

来源：

GENE | 1994年 / 146卷 / 02期

关键词：

GLUTATHIONE S-TRANSFERASE FUSION PROTEIN; AFFINITY CHROMATOGRAPHY; COUP-TF-LIKE FACTOR;

D O I：

10.1016/0378-1119(94)90307-7

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

We have produced a truncated form of the human estrogen receptor (hER) as a fusion protein with glutathione S-transferase (GST) in Spodoptera frugiperda (Sf) cells using the baculovirus expression vector (BEV) system. The protein is correctly produced and can be purified from crude whole-cell extracts by a single-step, batch-wise affinity-purification procedure. We show that this GST-hER fusion protein binds at its DNA-binding site specifically and in a hormone-inducible manner. Furthermore, we used the purified hER to analyze the complex estrogen response element (ERE) in the promoter of the oxytocin-encoding gene.

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收藏

页码：285 / 289

页数：5

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