Bacteria/cytology/genetics; Cell Death; Cesium/pharmacology; Chlorides/pharmacology; DNA Topoisomerases, Type I/metabolism; DNA, Superhelical/chemistry/genetics/isolation & purification/metabolism; Electrophoresis, Agar Gel; Ethidium/metabolism/pharmacology; Genome, Bacterial/genetics; Humans; Intercalating Agents/metabolism/pharmacology; Nucleic Acid Amplification Techniques; Plasmids/genetics/isolation & purification
Abstract :
[en] Several biochemical and biophysical methods are available to study the intercalation of a small molecule between two consecutive base pairs of DNA. Among them, the topoisomerase I-mediated DNA relaxation assay has proved highly efficient, relatively easy to handle and very informative to investigate drug binding to DNA. The test relies on the use of a supercoiled plasmid to mimic the topological constraints of genomic DNA. The three main components of the assay - the topoisomerase I enzyme, DNA helix and intercalating small molecules - are presented here in a structural context. The principle of the assay is described in detail, along with a typical experimental protocol.
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