A High-Throughput Assay for DNA Topoisomerases and Other Enzymes, Based on DNA Triplex Formation View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

2009-11-06

AUTHORS

Matthew R. Burrell , Nicolas P. Burton , Anthony Maxwell

ABSTRACT

We have developed a rapid, high-throughput assay for measuring the catalytic activity (DNA supercoiling or relaxation) of topoisomerase enzymes that is also capable of monitoring the activity of other enzymes that alter the topology of DNA. The assay utilises intermolecular triplex formation to resolve supercoiled and relaxed forms of DNA, the principle being the greater efficiency of a negatively supercoiled plasmid to form an intermolecular triplex with an immobilised oligonucleotide than the relaxed form. The assay provides a number of advantages over the standard gel-based methods, including greater speed of analysis, reduced sample handling, better quantitation and improved reliability and accuracy of output data. The assay is performed in microtitre plates and can be adapted to high-throughput screening of libraries of potential inhibitors of topoisomerases including bacterial DNA gyrase. More... »

PAGES

257-266

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/978-1-60327-418-0_16

DOI

http://dx.doi.org/10.1007/978-1-60327-418-0_16

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1012245669

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/19997889


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