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色氨酸阻遏蛋白如何与其操纵基因结合。

How Trp repressor binds to its operator.

作者信息

Staacke D, Walter B, Kisters-Woike B, von Wilcken-Bergmann B, Müller-Hill B

机构信息

Institut für Genetik der Universität zu Köln, FRG.

出版信息

EMBO J. 1990 Jun;9(6):1963-7. doi: 10.1002/j.1460-2075.1990.tb08324.x.

Abstract

We propose that the generally accepted model of a single Trp repressor dimer binding to a center of symmetry in the natural trp operator (Otwinowski et al., 1988) is wrong. We show here that the Trp repressor binds to a sequence whose center is located four base pairs either to the right or to the left of the central axis of symmetry that was previously identified. We show that: (i) the oligonucleotide used by Otwinowski et al. is not retarded by the Trp repressor in a mobility shift assay under conditions wherein a shorter oligonucleotide carrying our consensus sequence is retarded, (ii) that methylation protection experiments on the full natural operator sequence and the short oligonucleotide protect similar patterns and (iii) that by varying every base in the shorter oligonucleotide, we can demonstrate an optimal sequence for Trp repressor binding.

摘要

我们认为,普遍接受的单个色氨酸阻遏物二聚体与天然色氨酸操纵基因(Otwinowski等人,1988年)中的对称中心结合的模型是错误的。我们在此表明,色氨酸阻遏物与一个序列结合,该序列的中心位于先前确定的对称中心轴右侧或左侧四个碱基对处。我们表明:(i)在迁移率变动分析中,当携带我们的共有序列的较短寡核苷酸被阻滞时,Otwinowski等人使用的寡核苷酸不会被色氨酸阻遏物阻滞,(ii)对完整天然操纵基因序列和短寡核苷酸进行的甲基化保护实验保护的模式相似,以及(iii)通过改变较短寡核苷酸中的每个碱基,我们可以证明色氨酸阻遏物结合的最佳序列。

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