用于观察负载下蛋白质-DNA 相互作用的力荧光装置。

4区生物学 Q3 Biochemistry, Genetics and Molecular Biology

Methods in enzymology Pub Date : 2024-01-01 Epub Date: 2024-01-16 DOI:10.1016/bs.mie.2024.01.003

Jaehun Jung, Subin Kim, Sang-Hyun Rah, Jayil Lee, Min Ju Shon

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引用次数: 0

摘要

本章探讨了研究蛋白质-DNA 相互作用的先进单分子技术，尤其侧重于使用力荧光装置研究复制蛋白 A (RPA)。它结合了磁镊子（MT）和全内反射荧光（TIRF）显微镜，能够详细观察 DNA 在机械应力下的行为。本章详细介绍了如何利用 DNA 发夹和裸 DNA 来研究 RPA 的结合动力学及其对 DNA 机械特性的影响。这种方法让人们更深入地了解了 RPA 在 DNA 复制、修复和重组中的作用，突出了它在维持基因组稳定性方面的重要意义。

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Force-fluorescence setup for observing protein-DNA interactions under load.

This chapter explores advanced single-molecule techniques for studying protein-DNA interactions, particularly focusing on Replication Protein A (RPA) using a force-fluorescence setup. It combines magnetic tweezers (MT) with total internal reflection fluorescence (TIRF) microscopy, enabling detailed observation of DNA behavior under mechanical stress. The chapter details the use of DNA hairpins and bare DNA to examine RPA's binding dynamics and its influence on DNA's mechanical properties. This approach provides deeper insights into RPA's role in DNA replication, repair, and recombination, highlighting its significance in maintaining genomic stability.

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来源期刊

Methods in enzymology 生物-生化研究方法

CiteScore

2.90

自引率

0.00%

发文量

308

审稿时长

3-6 weeks

期刊介绍： The critically acclaimed laboratory standard for almost 50 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Each volume is eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 500 volumes the series contains much material still relevant today and is truly an essential publication for researchers in all fields of life sciences, including microbiology, biochemistry, cancer research and genetics-just to name a few. Five of the 2013 Nobel Laureates have edited or contributed to volumes of MIE.