Author:
Date:
2020-12-05
[Abstract] DNA footprinting is a classic technique to investigate protein-DNA interactions. However, traditional footprinting protocols can be unsuccessful or difficult to interpret if the binding of the protein to the DNA is weak, the protein has a fast off-rate, or if several different protein-DNA complexes are formed. Our protocol differs from traditional footprinting protocols, because it provides a method to isolate the protein-DNA complex from a native gel after treatment with the footprinting agent, thus removing the bound DNA from the free DNA or other protein-DNA complexes. The DNA is then ...
[摘要] [摘要] DNA足迹是研究蛋白质-DNA相互作用的经典技术。但是,如果蛋白质与DNA的结合较弱,蛋白质的脱落速率较快,或者形成了几种不同的蛋白质-DNA复合物,则传统的足迹方案可能会失败或难以解释。我们的协议不同于传统的足迹协议,因为它提供了一种在使用足迹剂处理后从天然凝胶中分离蛋白质-DNA复合物的方法,从而从游离DNA或其他蛋白质-DNA复合物中去除了结合的DNA。然后从分离的复合物中提取DNA,然后在测序凝胶上电泳以确定印迹模式。该分析为无法使用传统足迹法确定蛋白质与DNA接触的人提供了可能的解决方案。
[背景]核酸酶/化学足迹是一个典型的方法来探测蛋白质-DNA相互作用(腊士和施米茨,1978;萨瑟-德怀特和Gralla,1989;汉普等人,2007) ...
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