Mycobacteria Protocols / Edition 2

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A collection of cutting-edge techniques for anyone interested in studying the molecular biology of mycobacteria, a medically significant organism responsible for both human and animal disease, including tuberculosis and leprosy. The book describes the basic techniques required for any study of mycobacteria, as well as providing methods for introducing DNA, for creating mycobacterial mutants, for obtaining defined gene knockouts by homologous recombination in both fast and slow growing species, for gene regulation and expression, for diagnostic purposes in detecting drug resistance, and for culture.

The book contains black-and-white illustrations.

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Editorial Reviews

Doody's Review Service
Reviewer: Rebecca T. Horvat, PhD, D (ABMM)(University of Kansas Medical Center)
Description: Noting that the mycobacteria are "difficult organisms to work with," the editors provide this book with details on laboratory protocols that will be valuable to scientists working in this area. This first edition was published in 1998, and much has changed in the tools available to researchers. Thus, this book is needed to promote and enhance research in this area.
Purpose: This book will be a useful reference for scientists working with mycobacteria. These new techniques and ideas are valuable to laboratory researchers since many of these methods may take weeks or months to complete and involve expensive technologies. The book will be invaluable to these scientists.
Audience: Scientists with expertise in the study of mycobacteria are the intended audience, but the book also would be helpful for graduate students or postdoctoral fellows working at the bench. The authors include both senior scientists and bench scientists.
Features: The techniques include advice on the isolation of DNA and RNA from the thick-walled mycobacteria. The book includes methods for the isolation and study of the cell wall lipids and well as the investigation of global gene expression using microarrays and the introduction of plasmids using electroporation. Very detailed descriptions include a diagram of a continuous cell culture system for mycobacteria. This system allows for the investigation of how this bacterium responds to changes in environment. An additional system shows how to study mycobacteria inside the amoeba.
Assessment: This will be a very valuable resource for scientists working in the field of mycobacteria. It updates the previous edition with the techniques that have been developed in the last 10 years.
Charles E. Edmiston
This book was conceived as a laboratory resource describing in concise detail current molecular techniques used in basic or applied studies involving the mycobacteria. The editors have collected in a single volume numerous molecular protocols, which have been contributed by an outstanding group of international investigators. The spectrum of protocols included encompass laboratory techniques for studying the molecular biology of the mycobacteria to procedures appropriate for diagnostic or epidemiologic studies. This text should be viewed as a valuable resource for any mycobacterial investigator engaged in basic science or clinical studies. It would also be extremely helpful to young investigators or students studying the molecular genetics or microbial pathogenesis of the medically important mycobacteria. The text has been organized into 31 separate chapters. The first third of the book presents several chapters on basic techniques for studying the mycobacteria such as cultivation and preparation of nucleic acid, extraction of RNA, preparation of proteins, and cell-wall fractions. There are several chapters on phage application, transposon mutagenesis, gene replacement, and other techniques for studying gene regulation and expression. The last seven chapters address the use of molecular techniques for laboratory diagnosis and detection of drug resistance. The protocols in each chapter are well documented with concise material and methods sections. The editors have included a helpful "notes" section at the end of each chapter written by the contributors describing their own experiences when using these techniques, including valuable suggestions for overcoming common laboratorypitfalls. This is an excellent resource for either the established or novice investigator. The editors have solicited contributions from over 50 experts, many of whom are recognized as international leaders in the molecular biology and pathogenesis of the mycobacteria. The breadth of techniques covered and the thorough documentation included makes this text an ideal acquisition for either a personal or institutional holding.
Presents the latest molecular biology techniques for studying mycobacteria, with step-by-step instructions ensuring reproducibility. Methods range from basic culture techniques, fractionation, and nucleic acid isolation to more advanced protocols for the introduction of cloned DNA into mycobacteria, the use of reporter genes, and the expression of foreign genes. Other techniques covered include mutagenesis, genome analysis, speciation, and RFLP analysis. Includes an introduction to using the mycobacterial database on the Web. Annotation c. by Book News, Inc., Portland, Or.

3 Stars from Doody
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Product Details

  • ISBN-13: 9781588298898
  • Publisher: Springer-Verlag New York, LLC
  • Publication date: 9/28/2008
  • Series: Methods in Molecular Biology Series, #465
  • Edition description: 2nd ed. 2009
  • Edition number: 2
  • Pages: 456
  • Product dimensions: 6.10 (w) x 9.30 (h) x 1.00 (d)

Table of Contents

1. Whole Genome Sequencing for Comparative Genomics and De Novo Genome Assembly

Andrej Benjak, Claudia Sala, and Ruben C. Hartkoorn

2. Whole Transcriptome Sequencing for High-resolution Transcriptomic Analysis in Mycobacterium tuberculosis

Andrej Benjak, Claudia Sala, and Ruben C. Hartkoorn

3. RNA Sequencing for Transcript 5’-end Mapping in Mycobacteria

Scarlet S. Shell, Michael R. Chase, Thomas R. Ioerger, and Sarah M. Fortune

4. Fractionation and Analysis of Mycobacterial Proteins

Megan C. Lucas, Lisa M. Wolfe, Rachel M. Hazenfield, Jade Kurihara, Nicole A. Kruh-Garcia, John Belisle, and Karen M. Dobos

5. Lipid and Lipoarabinomannan Isolation and Characterization

Marie-Antoinette Lanéelle, Jérôme Nigou, and Mamadou Daffé

6. Metabolomics of Mycobacterium tuberculosis

Madhumitha Nandakumar, Gareth A. Prosser, Luis Pedro S. de Carvalho, and Kyu Rhee

7. Electroporation of Mycobacteria

Renan Goude, David M. Roberts, and Tanya Parish

8. Targeted Gene Knockout and Essentiality Testing by Homologous Recombination

Krishnamoorthy Gopinath, Digby F. Warner, and Valerie Mizrahi

9. Construction of Conditional Knockdown Mutants in Mycobacteria

Dirk Schnappinger, Kathryn M. O’Brien, and Sabine Ehrt

10. Mycobacterial Recombineering

Kenan C. Murphy, Kadamba Papavinasasundaram, and Christopher M. Sassetti

11. In vitro Models that Utilize Hypoxia to Induce Non-replicating Persistence in Mycobacteria

Charles D. Sohaskey and Martin I. Voskuil

12. Genetic Dissection of Mycobacterial Biofilms

Anil K Ojha, William R. Jacobs Jr., and Graham F. Hatfull

13. Measuring Efflux and Permeability in Mycobacteria

Liliana Rodrigues, Miguel Viveiros, and José A. Aínsa

14. Single-cell Analysis of Mycobacteria using Microfluidics and Time-lapse Microscopy

Neeraj Dhar and Giulia Manina

15. Antimicrobial Susceptibility Testing for Mycobacterium sp.

Delia Blanco-Ruano, David M. Roberts, Ruben Gonzalez-Del-Rio, Daniel Álvarez, Mª José Rebollo, Esther Pérez-Herrán, and Alfonso Mendoza

16. Determination of Compound Kill Kinetics Against Mycobacterium tuberculosis

Julie Early and Torey Alling

17. Microplate Alamar Blue Assay (MABA) and Low Oxygen Recovery Assay (LORA) for Mycobacterium tuberculosis

Sanghyun Cho, Hyung Sup Lee, and Scott Franzblau

18. A Multi-Stress Model for High Throughput Screening Against Non-replicating Mycobacterium tuberculosis

Ben Gold, Thulasi Warrier, and Carl Nathan

19. Isolation and Characterization of Compound-resistant Isolates of Mycobacterium tuberculosis

Theresa O’Malley and Eduard Melief

20. Macrophage Infection Models for Mycobacterium tuberculosis

Benjamin K. Johnson and Robert B. Abramovitch

21. Infection of Human Neutrophils to Study Virulence Properties of Mycobacterium tuberculosis

Tobias Dallenga, Björn Corleis, and Ulrich E. Schaible

22. Isolation of Bead-phagosomes to Study Virulence Function of M. tuberculosis Cell Wall Lipids

Anna Christina Geffken, Emmanuel C. Patin, and Ulrich E. Schaible

23. Live Imaging of Mycobacterium marinum Infection in Dictyostelium discoideum

Caroline Barisch, Ana T. López-Jiménez, and Thierry Soldati

24. Testing Chemical and Genetic Modulators in Mycobacterium tuberculosis Infected Cells using Phenotypic Assays

Vincent Delorme, Ok-Ryul Song, Alain Baulard, and Priscille Brodin

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