Table of Contents

Assaying DNA Topoisomerase I Relaxation Activity.- DNA Topoisomerase II-Catalyzed DNA Decatenation.- Plasmid DNA Supercoiling by DNA Gyrase.- Analyzing Reverse Gyrase Activity.- Topoisomerase II-Catalyzed ATP Hydrolysis as Monitored by Thin-Layer Chromatography.- Use of a Real-Time, Coupled Assay to Measure the ATPase Activity of DNA Topoisomerase II.- Analysis of Topoisomerase-DNA Interactions by Electrophoretic Mobility Shift Assay.- Filter Binding Assays for Topoisomerase—DNA Complexes.- DNATopoisomerase I-Mediated Nicking of Circular Duplex DNA.- Analysis of Cleavage Complexes Using Reactive Inhibitor Derivatives.- Uncoupling of Topoisomerase-Mediated DNA Cleavage and Religation.- Synthesis and Use of DNA Containing a 5?-Bridging Phosphorothioate as a Suicide Substrate for Type I DNA Topoisomerases.- Isolation of Covalent Enzyme-DNA Complexes.- ICE Bioassay.- DNA-Unwinding Test Using Eukaryotic DNA Topoisomerase I.- Drug—DNA Interactions.- Quinolone Interactions with DNA and DNA Gyrase.- Bactericidal Assays for Fluoroquinolones.- Drug Toxicity in E. coli Cells Expressing Human Topoisomerase I.- Drug-Induced Cytotoxicity in Tissue Culture.- Analysis of Drug Uptake-Flow Cytometry.- Cell-Cycle Analysis of Drug-Treated Cells.- Assaying Drug-Induced Apoptosis.- Single-Strand Conformational Polymorphism Analysis of DNA Topoisomerases I and II.- PCR-Based Cloning of DNA Topoisomerase Genes.- Topoisomerase II-Catalyzed Relaxation and Catenation of Plasmid DNA.- Topoisomerase II-Mediated Cleavage of Plasmid DNA.- Drug-induced Stabilization of Covalent DNA Topoisomerase I-DNA Intermediates.- Studying DNATopoisomerase I-Targeted Drugs In the Yeast.- Yeast Systems for Demonstrating the Targets of Anti-Topoisomerase II Agents.

Reviews

"This book completes a comprehensive collection of assays, including the weird and wonderful, that can be used to investigate the enzymology and drug inhibition of topoisomerases...I recommend any students about to undertake a PhD degree in this area to buy a copy. Even those who are well versed in the biology of topoisomerases should consider buying both volumes of this book, they will surely find some things they didn't know - I did!" - MicroBiology Today

"Described by expert experimentalists who have perfected the techniques, these reproducible methods include assays for enzyme-catalyzed DNA relaxation/supercoiling, DNA cleavage, DNA nicking, DNA decatentaion, and ATP hydrolysis." - Anticancer Research

"An unprecedented collection of cutting-edge experimental protocols for investigating the catalytic activities of DNA topoisomerases, as well as their specific interactions with topoisomerase-targeted antitumor and antibacterial drugs." -Tumori