High-Throughput Microtitre Plate-Based Assay for DNA Topoisomerases View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

2011-11-04

AUTHORS

James A. Taylor , Nicolas P. Burton , Anthony Maxwell

ABSTRACT

We have developed a rapid, high-throughput assay for measuring the catalytic activity (DNA supercoiling or relaxation) of DNA topoisomerases. The assay utilizes intermolecular triplex formation between an immobilized triplex-forming oligo (TFO) and a triplex-forming region inserted into the plasmid substrate (pNO1), and capitalizes on the observation that supercoiled DNA forms triplexes more readily than relaxed DNA. Thus, supercoiled DNA is preferentially retained by the TFO under triplex-forming conditions while relaxed DNA can be washed away. Due to its high speed of sample analysis and reduced sample handling over conventional gel-based techniques, this assay can be used to screen chemical libraries for novel inhibitors of topoisomerases. More... »

PAGES

229-239

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/978-1-61779-424-7_17

DOI

http://dx.doi.org/10.1007/978-1-61779-424-7_17

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1014413294

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/22130995


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