Kaitlin Lowran, Vereena Salib, Emma Cismas, Colin G Wu
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引用次数: 0
Abstract
Biolayer interferometry (BLI) is a powerful tool that enables direct observations of protein-G4 interactions in real-time. In this article, we discuss the crucial aspects in conducting a BLI experiment by using the TAR DNA-binding protein (TDP43) and a G4 DNA formed by (GGGGCC)4 as a sample application. We also describe the necessary precautions in designing the DNA substrate and evaluating the signal contributions arising from nonspecific binding interactions. A comprehensive guide is included that details the necessary materials and reagents, experimental procedures, and data analysis methods for researchers who are interested in using BLI for similar studies. The insights provided in this article will allow researchers to harness the potential of BLI and unravel the complexities of protein-G4 interactions with precision and confidence.
生物层干涉测量法（Biolayer interferometry，BLI）是一种强大的工具，可直接实时观测蛋白质与 G4 的相互作用。在本文中，我们以 TAR DNA 结合蛋白（TDP43）和由 (GGGGCC)4 形成的 G4 DNA 为应用样本，讨论了进行 BLI 实验的关键环节。我们还介绍了设计 DNA 底物和评估非特异性结合相互作用产生的信号贡献的必要注意事项。本文还附有一份综合指南，详细介绍了必要的材料和试剂、实验步骤以及数据分析方法，供有兴趣使用 BLI 进行类似研究的研究人员参考。本文提供的见解将使研究人员能够利用 BLI 的潜力，精确而自信地揭示蛋白质-G4 相互作用的复杂性。
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.
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