Mycobacterium Tuberculosis Protocols 1st Edition by Tanya Parish, Neil Stoker ISBN 978-1592591473 1592591473

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Product details: 

ISBN 10:1592591473

ISBN 13: 978-1592591473

Author: Tanya Parish, Neil G. Stoker

The aim of this book is to provide detailed protocols for studying the molecular biology of the pathogen Mycobacterium tuberculosis, and its int- actions with host cells. As established mycobacterial laboratories move – wards exploiting the genome, and laboratories with expertise in other fields apply them to mycobacteria, both traditional and novel methodologies need to be reviewed. Thus the chapters in Mycobacterium tuberculosis Protocols range from perspectives on storage of strains and safety issues to the application of the latest functional genomics technologies. The last few years have been remarkable ones for research into M. tuber- losis. The most important landmark by far has been the completion of the genome sequence of the widely studied H37Rv strain (1). We can now predict every protein and RNA molecule made by the pathogen. This information is or will soon be enriched by the addition of genome sequences of other strains from the M. tuberculosis complex: a second strain of M. tuberculosis, My- bacterium bovis, and the vaccine strain, M. bovis BCG. Valuable comparative data will also be provided by the genome sequences of Mycobacterium leprae, Mycobacterium avium, and Streptomyces coelicolor. Another recent milestone for M. tuberculosis has been the development of efficient mutagenesis me- odologies, the lack of which has been a major handicap in functional studies.

Table of contents: 

1. Plasmid Vectors
Nicola Casali, Sabine Ehrt

2. Isolation of DNA from Mycobacterium tuberculosis
Paul D. van Helden, Thomas C. Victor, Robin M. Warren, Eileen G. van Helden

3. Extraction of RNA from Intracellular Mycobacterium tuberculosis
Irene M. Monahan, Joseph A. Mangan, Philip D. Butcher

4. Transposon Mutagenesis in Mycobacteria Using Conditionally Replicating Mycobacteriophages
Stoyan S. Bardarov, Svetoslav S. Bardarov Jr., William R. Jacobs Jr.

5. Gene Replacement and Transposon Delivery Using the Negative Selection Marker sacB
Mary Jackson, Luis Reinaldo Camacho, Brigitte Gicquel, Christophe Guilhot

6. Gene Replacement Using Pretreated DNA
Bhavna G. Gordhan, Tanya Parish

7. Gene Replacement in Mycobacterium tuberculosis and Mycobacterium bovis BCG Using rpsL as a Dominant Negative Selectable Marker
Peter Sander, Burkhard Springer, Erik C. Böttger

8. Transcription Start-Site Mapping
Farahnaz Movahedzadeh, Jorge A. Gonzalez-Y-Merchand, Robert A. Cox

9. Fluorescence and Brightfield Cytology of Live Mycobacterium tuberculosis Cells
Rory P. Cooney, Natalie J. Garton, Michael R. Barer

10. Phage Replication Technology for Diagnosis and Drug Susceptibility Testing
Ruth McNerney

11. Detection of Mutations in Mycobacterium tuberculosis by a Dot Blot Hybridization Strategy
Thomas C. Victor, Paul D. van Helden

12. Restriction Fragment Length Polymorphism Typing of Mycobacteria
Dick van Soolingen, Petra E. W. de Haas, Kristin Kremer

13. Preparation of Culture Filtrate Proteins from Mycobacterium tuberculosis
Ida Rosenkrands, Peter Andersen

14. Analysis of the Capsule of Mycobacterium tuberculosis
Mamadou Daffé, Marie-Antoinette Lanée

15. Analysis of the Lipids of Mycobacterium tuberculosis
Richard A. Slayden, Clifton E. Barry 3rd

16. In Vitro Model of Hypoxically Induced Nonreplicating Persistence of Mycobacterium tuberculosis
Lawrence G. Wayne

17. Macrophage Virulence Assays
Pauline T. Lukey, Elizabeth U. Hooker

18. Analysis of Mycobacterium-Infected Macrophages by Immunoelectron Microscopy and Cell Fractionation
Wandy Beatty, David G. Russell

19. Real-Time PCR Using Molecular Beacons
Riccardo Manganelli, Sanjay Tyagi, Issar Smith

20. Electronic Access to Mycobacterium tuberculosis Sequence Data
Julian Parkhill

21. Proteomics
Joanna C. Betts, Marjorie A. Smith

22. Functional Genomics of Mycobacterium tuberculosis Using DNA Microarrays
Michael Wilson, Martin Voskuil, Dirk Schnappinger, Gary K. Schoolnik

23. Storage of Mycobacterial Strains
Kristin Kremer, Tridia van der Laan, Dick van Soolingen

24. Safety in the Laboratory
Heidi Alderton, Debbie Smith

25. Back Matter

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