DNA Topoisomerase Protocols: Volume I: DNA Topology and Enzymes / Edition 1

by Mary-Ann Bjornsti, Neil Osheroff

Overview

Beginning with the Escherichia coli co protein, or bacterial DNA topoisomerase I, an ever-increasing number of enzymes has been identified that catalyze changes in the linkage of DNA strands. DNA topoisomerases are ubiquitous in nature and have been shown to play critical roles in most processes involving DNA, including DNA replication, transcription, and rec- bination. These enzymes further constitute the cellular targets of a number of clinically important antibacterial and anticancer agents. Thus, further studies of DNA topology and DNA topoisomerases are critical to advance our und- standing of the basic biological processes required for cell cycle progression, cell division, genomic stability, and development. In addition, these studies will continue to provide critical insights into the cytofoxic action of drugs that target DNA topoisomerases. Such mechanistic studies have already played an important role in the development and clinical application of antimicrobial and chemotherapeutic agents. The two volumes of DNA Topoisomerase Prools are designed to help new and established researchers investigate all aspects of DNA topology and the function of these enzymes. The chapters are written by prominent investigators in the field and provide detailed background information and st- by-step experimental prools. The topics covered in Volume I; DNA Topology and Enzymes, range from detailed methods to analyze various aspects of DNA structure, from linking number, knotting/unknotting, site-specific recombi- tion, and decatenation to the overexpression and purification of bacterial and eukaryotic DNA topoisomerases from a variety of cell systems and tissues.

Product Details

ISBN-13:	9781617370397
Publisher:	Springer-Verlag New York, LLC
Publication date:	11/10/2010
Series:	Methods in Molecular Biology , #94
Edition description:	Softcover reprint of hardcover 1st ed. 1999
Pages:	327
Product dimensions:	5.98(w) x 9.02(h) x 0.03(d)

Table of Contents

to DNA Topoisomerases.- Resolution of DNA Molecules by One-Dimensional Agarose-Gel Electrophoresis.- Two-Dimensional Agarose-Gel Electrophoresis of DNA Topoisomers.- Analysis of Altered DNA Structures.- Purification of Supercoiled Plasmid DNA.- Purification and Use of DNA Minicircles with Different Linking Numbers.- Isolation of Kinetoplast DNA.- Isolation of Knotted DNA from Coliphage P4.- Analysis of DNA Knots and Catenanes by Agarose-Gel Electrophoresis.- Sedimentation Analysis of Bacterial Nucleoid Structure.- Coating DNA with RecA Protein to Distinguish DNA Path by Electron Microscopy.- Methods for Analyzing DNA Bending.- Formation of Extrachromosomal DNA Rings in Saccharomyces cerevisiae Using Site-Specific Recombination.- Overexpression and Purification of Bacterial DNA Gyrase.- Overexpression and Purification of Bacterial DNA Topoisomerase I.- Overexpression and Purification of Escherichia coliDNA Topoisomerase III.- Overexpression and Purification of Bacterial Topoisomerase IV.- Purification of the Bacteriophage T4 Type II DNA Topoisomerase.- Overexpression and Purification of DNA Topoisomerase I from Yeast.- Overexpression and Purification of Saccharomyces cerevisiae DNA Topoisomerase II from Yeast.- Purification of DNA Topoisomerase II from Drosophila melanogaster.- Purification of DNA Topoisomerase I from Human Placenta.- Purification of Baculovirus-Expressed Human DNA Topoisomerase I.- Rapid Purification of DNA Topoisomerase II Containing a Hexahistidine Tag by Metal Ion Affinity Chromatography.- Metabolic Labeling, Immunoprecipitation, and Two-Dimensional Tryptic Phosphopeptide Mapping of Human Topoisomerase II.- Immunoblot Analysis and Band Depletion Assays.- Visualization of DNA Topoisomerases by Electron Microscopy.- Appendix: Compendium of DNA Topoisomerase Sequences.

Editorial Reviews

"...an invaluable practical aid. The central and expanding role of these enzymes in basic cell biology together with their important pharmaceutical applications will ensure that book will remain relevant for some years to come."-Cell Biology International 2000

"The introductory chapter by the editors gives a beautifully brief and concise overview of the entire family of DNA topoisomerases and related enzymes and can be used as an Ariadne's thread to the key papers of the field. Another useful item is the appended compendium of topoisomerase sequences, which is likely to be helpful in designing expression vectors, deletion mutants, etc. Most of the other chapters contain a lot of useful information and easily followed procedures.a very valuable addition to the topoisomerase literature. It summarizes a lot of useful information for those engaged or getting engaged in research on DNA topology or topoisomerases. Those just wanting to carry out some of the analytical procedures will in most cases find what they need, i.e. a concise delineation of the analytical background, some detailed and easy to follow prools, and a comprehensive guideline for the interpretation of the results. As such, the book definitely deserves a place on many laboratory shelves"-FEBS Letters

"The introductory chapter is one of the most concise and clearly presented summaries of our current knowledge that has been written, and it will be useful even for experts in the field....The volume concludes with a well-organized and useful compendium of toposomerase DNA sequences. Volume I is clearly a valuable source of methodological expertise for anyone studying topoisomerases."-New Biological Books

"...We recommend greatly this remarkable book to all our readers, without exception." - Cellular and Molecular Biology

From the Publisher