一种测量嵌入剂解旋角度和环状DNA超螺旋密度的快速方法。
A rapid method for the measurement of the unwinding angle of intercalating agents and the superhelix density of circular DNAs.
作者信息
Lee J S, Morgan A R
出版信息
Nucleic Acids Res. 1978 Jul;5(7):2425-39. doi: 10.1093/nar/5.7.2425.
Abstract
By incubating covalently-closed circular DNA in the presence of calf thymus topoisomerase and unwinding ligands (intercalating drugs and proteins) DNAs of different superhelix density can be produced. These changes in superhelix density can be monitored by an ethidium fluorescence assay, since the level of ethidium binding varies with the superhelix density of a DNA. Thus the equivalence point, in a titration between an unwinding ligand and a supercoiled DNA, can be found. This forms the basis for an extremely rapid method for measuring unwinding angles and superhelix densities. Results are presented which agree well with those reported by previous authors using different techniques. The present method compares very favourably with others when evaluated in terms of rapidity, cost of materials, cost of equipment, accuracy and also applicability.
摘要
通过在小牛胸腺拓扑异构酶和解旋配体（嵌入药物和蛋白质）存在的情况下孵育共价闭合环状DNA，可以产生不同超螺旋密度的DNA。超螺旋密度的这些变化可以通过溴化乙锭荧光测定法进行监测，因为溴化乙锭的结合水平随DNA的超螺旋密度而变化。因此，可以找到解旋配体与超螺旋DNA滴定中的等效点。这构成了一种测量解旋角度和超螺旋密度的极其快速的方法的基础。所呈现的结果与先前作者使用不同技术报道的结果非常吻合。在速度、材料成本、设备成本、准确性以及适用性方面进行评估时，本方法与其他方法相比具有很大优势。
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