利用 DNA 环化实验研究鱼腥藻感应视紫红质转导蛋白（ASRT）介导的 DNA 环化动力学。

Kinetics of DNA looping by Anabaena sensory rhodopsin transducer (ASRT) by using DNA cyclization assay.

机构信息

Department of Physics, Sogang University, Seoul, Korea.

Department of BioNanoScience Kavli Institute of Nanoscience of Delft University of Technology, Delft, The Netherlands.

出版信息

Sci Rep. 2021 Dec 9;11(1):23721. doi: 10.1038/s41598-021-03148-4.

DOI:10.1038/s41598-021-03148-4

PMID:34887464

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8660804/

Abstract

DNA cyclization assay together with single-molecule FRET was employed to monitor protein-mediated bending of a short dsDNA (~ 100 bp). This method provides a simple and easy way to monitor the structural change of DNA in real-time without necessitating prior knowledge of the molecular structures for the optimal dye-labeling. This assay was applied to study how Anabaena sensory rhodopsin transducer (ASRT) facilitates loop formation of DNA as a possible mechanism for gene regulation. The ASRT-induced DNA looping was maximized at 50 mM of Na, while Mg also played an essential role in the loop formation.

摘要

采用 DNA 环化分析与单分子Förster 共振能量转移（FRET）相结合的方法，实时监测蛋白介导的短双链 DNA（~100bp）弯曲。该方法无需预先了解最佳荧光染料标记的分子结构，提供了一种简单易用的监测 DNA 结构变化的方法。该方法用于研究蓝藻感应视紫红质转导蛋白（ASRT）如何促进 DNA 环化形成，这可能是一种基因调控的机制。ASRT 诱导的 DNA 环化在 50mM 的 Na 下达到最大值，而 Mg 也在环化形成中起关键作用。