Quantifying Single Molecule DNA-ligand Interactions

本项目的目标是开发单分子操作方法，用于定量研究DNA-配体相互作用，并使用这些方法来研究特定的生物学重要的相互作用。除了方法的发展，这项工作将揭示重要的问题，从DNA-小分子相互作用的基本生物物理学的生物物理和生物化学机制的复杂蛋白质相互作用参与DNA复制和重组。 为实现上述目标，将致力于以下三个具体目标。1.利用DNA拉伸技术探讨DNA与小分子相互作用的生物物理机制。2. 探讨单链DNA结合蛋白的结合机制，单链DNA结合蛋白是DNA复制的重要组成部分。3.研究噬菌体T4系统中重要的DNA复制和重组蛋白的作用。PI开发了创新的生物物理方法，使用单分子操作来检查DNA结合的细节。这些结果将对该领域以外的领域产生重大影响;因为这些方法可能适用于检查各种重要的生物系统。PI还在物理，化学和生物学的跨学科研究中培养研究生和本科生，包括与少数民族服务和本科院校的积极合作。这项研究的更广泛的影响也延伸到课堂上，PI开发了一个新的高级物理实验室课程，其中包含几个先进的研究水平的实验，包括使用光镊和原子力显微镜。PI开发的实验公开提供给其他机构的科学家使用。PI还为本科教学做出了重大贡献，将他的研究成果带入课堂，如药学物理和生命科学物理。该项目由生物科学理事会分子和细胞生物科学部的分子生物物理学计划以及数学和物理科学理事会的物理学部支持。

The objective of this project is to develop single molecule manipulation methods for the quantitative study of DNA-ligand interactions and to use these methods to investigate specific biologically important interactions. In addition to method development, this work will shed light on important problems ranging from the fundamental biophysics of DNA-small molecule interactions to the biophysical and biochemical mechanisms of complex protein interactions involved in DNA replication and recombination. The following three specific aims will be addressed to accomplish the above goals. 1. To probe the biophysical mechanism of DNA-small molecule interactions using DNA stretching. 2. To probe the binding mechanism of single-stranded DNA binding proteins, which are essential components of DNA replication. 3. To investigate the role of important DNA replication and recombination proteins from the bacteriophage T4 system. The PI has developed innovative biophysical methods for examining DNA binding in exquisite detail using single molecule manipulation. The results will have a significant impact beyond this field; as such methods may be adapted to examine a wide variety of important biological systems. The PI also has a strong track record of training graduate and undergraduate students in interdisciplinary research at the interface of physics, chemistry, and biology, including active collaborations with minority-serving and undergraduate institutions. The broader impacts of this research also extend into the classroom, where the PI has developed a new Advanced Physics Laboratory course, which contains several advanced research level experiments, including the use of optical tweezers and atomic force microscopy. Experiments developed by the PI are made publicly available for use by scientists at other institutions. The PI also makes significant contributions to undergraduate teaching, bringing examples from his research into the classroom in courses such as Physics for Pharmacy and Physics for the Life Sciences. This project is supported by the Molecular Biophysics Program in the Division of Molecular and Cellular Biosciences in the Directorate for Biological Sciences and the Division of Physics in the Mathematical and Physical Sciences Directorate.