DNA Topoisomerase Protocols, I: DNA Topology and Enzymes brings together an unprecedented collection of cutting-edge experimental protocols for investigating DNA structure, topology, and DNA topoisomerase function. Described by expert experimentalists who have perfected the techniques, these unfailingly reproducible methods utilize new approaches to study changes in DNA topology (linking number, knotting, catenation, and relaxation) and DNA structure (bending), as well as to assess chromosome structure through FLP-mediated recombination and to analyze bacterial nucleoid structure. State-of-the-art protocols also detail the expression and purification of bacterial, viral, and eukaryotic DNA topoisomerases from a variety of sources, including baculovirus, and bacterial, yeast and mammalian expression systems. A companion volume, DNA Topoisomerase Protocols, v. 2: Enzymology and Drugs, provides detailed protocols to study the catalytic activities of DNA topoisomerases, as well as their specific interactions with topoisomerase-targeted antitumor and antibacterial drugs. DNA Topoisomerase Protocols, I: DNA Topology and Enzymes will be of immense value to the many basic scientists and clinicians who want better to understand and to exploit proficiently the wealth of recent discoveries about chromatin structure and its relation to gene expression, about DNA topoisomerases as the targets of antitumor and antibacterial agents, and about DNA repair.

Beginning with the Escherichia coli ? protein, or bacterial DNA topoisomerase I, an ever-increasing number of enzymes have 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 p- cesses 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 cytotoxic 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 Protocols 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 protocols. The topics covered in Part 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.

The problem of unraveling two intertwined strands during the duplication of DNA was recognized shortly after the proposal of the DNA double helix structure in 1953. A group of enzymes called DNA topoisomerases solve this problem by breaking and rejoining DNA molecules in a controlled manner, thereby allowing strands to be passed through each other and thus untangled—not just during DNA replication, but also during many other basic cellular processes. Because of their intimate involvement in the workings of the cell, topoisomerases are also the logical targets of many antibiotics (including Cipro) and anticancer agents. This book, written by James Wang, the discoverer of the first topoisomerase and a leader in the field since, presents ten chapters covering the historical backdrop of the DNA entanglement problem and the discovery of the DNA topoisomerases, how DNA topoisomerases perform their magic in DNA replication, transcription, genetic recombination and chromosome condensation, and how they are targets of therapeutic agents. The book should appeal to readers from undergraduates upwards with interests in the biological and clinical aspects of topoisomerase function, or in the mathematics and physics of topology.

Each volume of Advances in Pharmacology provides a rich collection of reviews on timely topics. Emphasis is placed on the molecular basis of drug action, both applied and experimental.

Updated to include the newest drugs and those currently in development, this Fifth Edition is a comprehensive reference on the preclinical and clinical pharmacology of anticancer agents. Organized by drug class, the book provides the latest information on all drugs and biological agents—their mechanisms of action, interactions with other agents, toxicities, side effects, and mechanisms of resistance. The authors explain the rationale for use of drugs in specific schedules and combinations and offer guidelines for dose adjustment in particular situations. This edition's introduction includes timely information on general strategies for drug usage, the science of drug discovery and development, economic and regulatory aspects of cancer drug development, and principles of pharmacokinetics. Eight new chapters have been added and more than twenty have been significantly revised. A companion website includes the fully searchable text and an image bank.

"A key aspect of DNA is its ability to form a variety of structures, this book explains the origins and importance of such structures"--Provided by publisher.

This book examines specific techniques which can be used to explore new drug targets and the effectiveness of new antibiotics. By testing new antimicrobial agents and modified existing drugs, the most vulnerable cell processes, such as cell wall and membrane synthesis, DNA replication, RNA transcription and protein synthesis, can be better exploited. This in-depth volume, however, delves even deeper by identifying additional novel cellular targets for these new therapies. The book will provide laboratory investigators with the vital tools they need to test the antimicrobial potential of products and to curb the rise of so many infectious diseases.

This exceptional laboratory manual describes thirty-seven procedures most likely to be used in the next decade for molecular, biochemical, and cellular studies on Drosophila. They were selected after extensive consultation with the research community and rigorously edited for clarity, uniformity, and conciseness.The methods included permit investigation of chromosomes, cell biology, molecular biology, genomes, biochemistry, and development. Each protocol includes the basic information needed by novices, with sufficient detail to be valuable to experienced investigators. Each method is carefully introduced and illustrated with figures, tables, illustrations, and examples of the data obtainable. The book's appendices include key aspects of Drosophila biology, essential solutions, buffers, and recipes.An evolution of Michael Ashburner's 1989 classic Drosophila: A Laboratory Manual, this book is an essential addition to the personal library of Drosophila investigators and an incomparable resource for other research groups with goals likely to require fly-based technical approaches.

In this new edition, the editors have thoroughly updated and dramatically expanded the number of protocols to take advantage of the newest technologies used in all branches of research and clinical medicine today. These proven methods include real time PCR, SNP analysis, nested PCR, direct PCR, and long range PCR. Among the highlights are chapters on genome profiling by SAGE, differential display and chip technologies, the amplification of whole genome DNA by random degenerate oligonucleotide PCR, and the refinement of PCR methods for the analysis of fragmented DNA from fixed tissues. Each fully tested protocol is described in step-by-step detail by an established expert in the field and includes a background introduction outlining the principle behind the technique, equipment and reagent lists, tips on trouble shooting and avoiding known pitfalls, and, where needed, a discussion of the interpretation and use of results.

Now fully updated and considerably expanded, Glycoanalysis Protocols, 2nd ed., makes available to all protein scientists, and particularly those working with today's pharmaceuticals, the most advanced and reproducible glycoanalysis techniques currently in use. Developed by highly experienced carbohydrate chemists, biochemists, and physical chemists, these detailed, up-to-date, and proven analytical techniques cover the areas of glycoprotein macromolecular structural analysis, oligosaccharide profiling, lipid conjugate characterization, microorganism structure determination, and proteoglycan function. Special attention has been given to advanced analytical techniques in biotechnology during the production of recombinant glycoproteins and other therapeutics. Hailed as "indispensable" in its first edition, Glycoanalysis Protocols, 2nd ed., continues with vital, time-tested techniques addressing the needs of both biomedical researchers and protein macromolecular structural chemists. It will well serve all those starting work on the analysis of glycoproteins, as well as more experienced investigators seeking to augment their expertise.