一种用于测定溶液中游离蛋白质 - DNA 结合亲和力的毛细管电泳迁移率变动分析方法。
A capillary electrophoresis mobility shift assay for protein-DNA binding affinities free in solution.
作者信息
Foulds G J, Etzkorn F A
机构信息
Department of Chemistry, University of Virginia, McCormick Road, Charlottesville, VA 22901, USA.
出版信息
Nucleic Acids Res. 1998 Sep 15;26(18):4304-5. doi: 10.1093/nar/26.18.4304.
Abstract
Quantitative determination of dissociation constants for DNA-protein complexes will help clarify the molecular mechanisms of transcription, replication and DNA repair. A practical capillary electrophoresis mobility shift assay (CEMSA) for protein-DNA affinities free in solution is presented. The method is fast and simple, precise and general. The speed (<2 min separations) and simplicity derive from the use of an uncoated capillary with no gel matrix. The dissociation constant for GCNK58, a DNA-binding-region construct of the yeast transcription factor GCN4, binding to the AP1 DNA site was measured ( K d = 35 +/- 4 nM) to demonstrate the utility of the method.
摘要
DNA-蛋白质复合物解离常数的定量测定将有助于阐明转录、复制和DNA修复的分子机制。本文介绍了一种用于测定溶液中蛋白质-DNA亲和力的实用毛细管电泳迁移率变动分析方法（CEMSA）。该方法快速简便、精确通用。其速度（分离时间<2分钟）和简便性源于使用无凝胶基质的未涂层毛细管。通过测定酵母转录因子GCN4的DNA结合区域构建体GCNK58与AP1 DNA位点的结合解离常数（Kd = 35 +/- 4 nM）来证明该方法的实用性。
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