用于原位实时测量蛋白质诱导的 DNA 构象变化的平台。
Platform for in situ real-time measurement of protein-induced conformational changes of DNA.
机构信息
Department of Biomedical Engineering, Boston University, Boston, MA 02215, USA.
出版信息
Proc Natl Acad Sci U S A. 2010 Jan 26;107(4):1397-401. doi: 10.1073/pnas.0912182107. Epub 2010 Jan 4.
Abstract
A platform for in situ and real-time measurement of protein-induced conformational changes in dsDNA is presented. We combine electrical orientation of surface-bound dsDNA probes with an optical technique to measure the kinetics of DNA conformational changes. The sequence-specific Escherichia coli integration host factor is utilized to demonstrate protein-induced bending upon binding of integration host factor to dsDNA probes. The effects of probe surface density on binding/bending kinetics are investigated. The platform can accommodate individual spots of microarrayed dsDNA on individually controlled, lithographically designed electrodes, making it amenable for use as a high throughput assay.
摘要
本文提出了一种用于实时原位测量蛋白质诱导 dsDNA 构象变化的平台。我们将表面结合 dsDNA 探针的电定向与光学技术相结合，以测量 DNA 构象变化的动力学。利用序列特异性的大肠杆菌整合宿主因子（integration host factor）来证明整合宿主因子与 dsDNA 探针结合时引起的 DNA 弯曲。研究了探针表面密度对结合/弯曲动力学的影响。该平台可以容纳微阵列 dsDNA 的单个点，这些点位于单独控制的光刻设计电极上，使其适合用作高通量测定。
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