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使用声波传感器对表面结合DNA的大小和形状进行定量测定。

Quantitative determination of size and shape of surface-bound DNA using an acoustic wave sensor.

作者信息

Tsortos Achilleas, Papadakis George, Mitsakakis Konstantinos, Melzak Kathryn A, Gizeli Electra

机构信息

Institute of Molecular Biology and Biotechnology, Foundation for Research & Technology Hellas, Vassilika Vouton, 71110 Heraklion, Greece.

出版信息

Biophys J. 2008 Apr 1;94(7):2706-15. doi: 10.1529/biophysj.107.119271. Epub 2008 Jan 4.

DOI:10.1529/biophysj.107.119271
PMID:18178642
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2267124/
Abstract

DNA bending plays a significant role in many biological processes, such as gene regulation, DNA replication, and chromosomal packing. Understanding how such processes take place and how they can, in turn, be regulated by artificial agents for individual oriented therapies is of importance to both biology and medicine. In this work, we describe the application of an acoustic wave device for characterizing the conformation of DNA molecules tethered to the device surface via a biotin-neutravidin interaction. The acoustic energy dissipation per unit mass observed upon DNA binding is directly related to DNA intrinsic viscosity, providing quantitative information on the size and shape of the tethered molecules. The validity of the above approach was verified by showing that the predesigned geometries of model double-stranded and triple-helix DNA molecules could be quantitatively distinguished: the resolution of the acoustic measurements is sufficient to allow discrimination between same size DNA carrying a bent at different positions along the chain. Furthermore, the significance of this analysis to the study of biologically relevant systems is shown during the evaluation of DNA conformational change upon protein (histone) binding.

摘要

DNA弯曲在许多生物学过程中发挥着重要作用,如基因调控、DNA复制和染色体包装。了解这些过程如何发生,以及它们如何反过来被人工制剂调控以进行个体化治疗,对生物学和医学都很重要。在这项工作中,我们描述了一种声波装置的应用,该装置用于表征通过生物素-中性抗生物素蛋白相互作用 tethered 到装置表面的DNA分子的构象。DNA结合时观察到的单位质量的声能耗散与DNA特性粘度直接相关,提供了关于 tethered 分子大小和形状的定量信息。通过表明可以定量区分模型双链和三链螺旋DNA分子的预先设计的几何形状,验证了上述方法的有效性:声学测量的分辨率足以区分沿着链在不同位置弯曲的相同大小的DNA。此外,在评估蛋白质(组蛋白)结合后DNA构象变化的过程中,显示了这种分析对生物学相关系统研究的重要性。