染色质蛋白DNA桥接效率的定量测定

Quantitative Determination of DNA Bridging Efficiency of Chromatin Proteins.

作者信息

van der Valk Ramon A, Qin Liang, Moolenaar Geri F, Dame Remus T

机构信息

Leiden Institute of Chemistry, Leiden University, Leiden, The Netherlands.

Leiden Institute of Chemistry and Centre for Microbial Cell Biology, Leiden University, Leiden, The Netherlands.

出版信息

Methods Mol Biol. 2018;1837:199-209. doi: 10.1007/978-1-4939-8675-0_12.

DOI:10.1007/978-1-4939-8675-0_12

PMID:30109613

Abstract

DNA looping is important for genome organization in all domains of life. The basis of DNA loop formation is the bridging of two separate DNA double helices. Detecting DNA bridge formation generally involves the use of complex single-molecule techniques (atomic force microscopy, magnetic, or optical tweezers). Although DNA bridging can be qualitatively described, quantification of DNA bridging and bridging dynamics using these techniques is challenging. Here, we describe a novel biochemical assay capable of not only detecting DNA bridge formation, but also allowing for quantification of DNA bridging efficiency and the effects of physico-chemical conditions on DNA bridge formation.

摘要

DNA环化对于生命所有领域的基因组组织都很重要。DNA环形成的基础是两个独立的DNA双螺旋的桥接。检测DNA桥形成通常涉及使用复杂的单分子技术（原子力显微镜、磁力或光镊）。尽管DNA桥接可以定性描述，但使用这些技术对DNA桥接及其动力学进行定量具有挑战性。在这里，我们描述了一种新型生化测定法，它不仅能够检测DNA桥的形成，还能对DNA桥接效率以及物理化学条件对DNA桥形成的影响进行定量。