Biosensor platform for transcription factor DNA binding activity detection

B. Tomé, R. Rodrigues, G. Ferreira
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Abstract

The general objective of this project is to develop an acoustic biosensor platform to study protein-DNA binding and its application to the study of DNA transcription factors. An impedance analysis methodology enables the calculation of the acoustic energy dissipation per unit mass observed upon DNA binding, providing quantitative information on the size and shape of the tethered molecules. The rationale of the project rely on the fact that the conformational changes and bending of DNA upon protein binding increase the rigidity of DNA films immobilized at the surface of an acoustic sensor. As a result less acoustic energy is dissipated what is signaled by a decrease of the variation of the acoustic motional resistance. Such experimental approach, together with the associated mathematical signal processing and physical modeling, will provide quantitative information on sequence and conformation DNA sequences recognized by specific transcriptional factors as well as affinity and kinetic constants.
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转录因子DNA结合活性检测的生物传感器平台
本项目的总体目标是开发一个声学生物传感器平台来研究蛋白质-DNA结合及其在DNA转录因子研究中的应用。阻抗分析方法可以计算DNA结合时观察到的每单位质量的声能量耗散,从而提供有关拴链分子大小和形状的定量信息。该项目的基本原理依赖于这样一个事实,即蛋白质结合时DNA的构象变化和弯曲增加了固定在声学传感器表面的DNA膜的刚性。因此,较少的声能被耗散,这是由声运动阻力变化的减少所发出的信号。这种实验方法,连同相关的数学信号处理和物理建模,将提供由特定转录因子识别的序列和构象DNA序列的定量信息,以及亲和和动力学常数。
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