Mycobacterium Tuberculosis Protocols -

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Résumé :
With one-third of the world's population infected with Mycobacterium tuberculosis, over two million people a year dying from tuberculosis, and the appearance of multidrug-resistant strains, the need to understand the biology of M. tuberculosis, and so to develop new interventions, has become acute. In Mycobacterium tuberculosis Protocols, leading investigators with extensive practical knowledge and experience describe their best methods for studying this dangerous pathogen. Packed with step-by-step instructions to ensure successful results, these methods range from basic handling techniques to the application of functional genomics. These molecular techniques are suitable for research in genetics, biochemistry, microbiology, cell biology, epidemiology, and diagnostics, and are at the forefront of biological research as a whole, as well as in focused M. tuberculosis research. Highlights include methods for the basic safety and culture of M. tuberculosis, fractionation of the bacterium (nucleic acids, lipids, culture filtrate, and capsule), the analysis of gene expression (start-site mapping, real-time PCR, microarrays, and proteomics), the growth of the bacterium in macrophages and low oxygen, cytological analysis of the bacteria, and diagnostics. Highly practical and accessible, Mycobacterium tuberculosis Protocols utilizes advanced functional genomics and mutagenesis methodologies to provide both experimental and clinical investigators all the powerful techniques needed to illuminate the molecular biology of tuberculosis and its interactions with host cells, and so drive work on the wide variety of emerging therapeutic opportunities.

Sommaire:
Plasmid Vectors Nicola Casali and Sabine Ehrt Isolation of DNA from Mycobacterium tuberculosis Paul D. van Helden, Thomas C. Victor, Robin M. Warren, and Eileen G. van Helden Extraction of RNA from Intracellular Mycobacterium tuberculosis: Methods, Considerations, and Applications Irene M. Monahan, Joseph A. Mangan, and Philip D. Butcher Transposon Mutagenesis in Mycobacteria Using Conditionally Replicating Mycobacteriophages Stoyan S. Bardarov, Svetoslav S. Bardarov, Jr., and William R. Jacobs, Jr Gene Replacement and Transposon Delivery Using the Negative Selection Marker sacB Mary Jackson, Luis Reinaldo Camacho, Brigitte Gicquel, and Christophe Guilhot Gene Replacement Using Pretreated DNA Bhavna G. Gordhan and Tanya Parish Gene Replacement in Mycobacterium tuberculosis and Mycobacterium bovis BCG Using rpsL+ as a Dominant Negative Selectable Marker Peter Sander, Burkhard Springer, and Erik C. B?ttger Transcription Start-Site Mapping Farahnaz Movahedzadeh, Jorge A. Gonzalez-Y-Merchand, and Robert A. Cox Fluorescence and Brightfield Cytology of Live M. tuberculosis Cells Rory P. Cooney, Natalie J. Garton, and Michael R. Barer Phage Replication Technology for Diagnosis and Drug Susceptibility Testing Ruth McNerney Detection of Mutations in Mycobacterium tuberculosis by a Dot Blot Hybridization Strategy Thomas C. Victor and Paul D. van Helden Restriction Fragment Length Polymorphism Typing of Mycobacteria Dick van Soolingen, Petra E. W. de Haas, and Kristin Kremer Preparation of Culture Filtrate Proteins from Mycobacterium tuberculosis Ida Rosenkrands and Peter Andersen Analysis of the Capsule of Mycobacterium tuberculosis Mamadou Daff? and Marie-Antoinette Lan?elle Analysis of the Lipids of Mycobacterium tuberculosis Richard A. Slaydenand Clifton E. Barry, 3rd In Vitro Model of Hypoxically Induced Nonreplicating Persistence of Mycobacterium tuberculosis Lawrence G. Wayne Macrophage Virulence Assays Pauline T. Lukey and Elizabeth U. Hooker Analysis of Mycobacterium-Infected Macrophages by Immunoelectron Microscopy and Cell Fractionation Wandy Beatty and David G. Russell Real-Time PCR Using Molecular Beacons: A New Tool to Identify Point Mutations and to Analyze Gene Expression in Mycobacterium tuberculosis Riccardo Manganelli, Sanjay Tyagi, and Issar Smith Electronic Access to Mycobacterium tuberculosis Sequence Data Julian Parkhill Proteomics Joanna C. Betts and Marjorie A. Smith Functional Genomics of Mycobacterium tuberculosis Using DNA Microarrays Michael Wilson, Martin Voskuil, Dirk Schnappinger, and Gary K. Schoolnik Storage of Mycobacterial Strains Kristin Kremer, Tridia van der Laan, and Dick van Soolingen Safety in the Laboratory Heidi Alderton and Debbie Smith