Chang Jeong Won, Ngabonziza Daniel, Deweese Joseph
Department of Biological, Physical, and Human Sciences, Freed-Hardeman University, Henderson, TN, USA.
Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN, USA.
Methods Mol Biol. 2025;2928:109-114. doi: 10.1007/978-1-0716-4550-5_10.
Binding between DNA topoisomerase II and plasmid DNA can be measured using a variety of methods. The electrophoretic mobility shift assay (EMSA) is an efficient and inexpensive means for examining DNA affinity. This method relies on the principle that increasing concentrations of enzyme will result in more topoisomerase bound to DNA. As more binding occurs, the DNA migration in an agarose gel is decreased causing an upward "shift" in the DNA molecules when visualized in the gel. This shift can be compared across a range of concentrations to establish binding parameters under varying circumstances such as substrate type, the presence of a compound or drug, incubation time, or other conditions.
DNA拓扑异构酶II与质粒DNA之间的结合可以使用多种方法进行测量。电泳迁移率变动分析（EMSA）是一种检测DNA亲和力的高效且经济的方法。该方法基于这样的原理：酶浓度的增加会导致更多的拓扑异构酶与DNA结合。随着结合的增加，琼脂糖凝胶中DNA的迁移减少，在凝胶中观察时会导致DNA分子向上“迁移”。可以在一系列浓度范围内比较这种迁移，以确定在不同情况下（如底物类型、化合物或药物的存在、孵育时间或其他条件）的结合参数。
