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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

Table of Contents Isolation of Mycobacterium ssp. Genomic DNA. John T. Belisle*, Spencer B. Mahaffey and Preston J. Hill. Isolation of Mycobacterial RNA. Tige R. Rustad, David M. Roberts, Reiling P. Liao and David R. Sherman* Lipoglycans of Mycobacterium tuberculosis: isolation, purification and characterization. Libin Shi, Jordi B. Torrelles and Delphi Chatterjee* Analysing lipid metabolism: activation and ß-oxidation of fatty acids.Paul Robert Wheeler* Analysis of lipid biosynthesis and location. Paul Robert Wheeler* Whole genome analysis using microarrays. S. J. Waddell,* J. Hinds and P. D. Butcher Use of DNA microarrays to study global patterns of gene expression. Roberta Provvedi, Giorgio Palù, Riccardo Manganelli* Two-dimensional gel electrophoresis-based proteomics of mycobacteria. Jens Mattow*, Frank Siejak, Kristine Hagens, Julia Kreuzeder, Stefan H.E. Kaufmann, Ulrich E. Schaible Transport assays and permeability in pathogenic mycobacteria. Marie-Antoinette Lanéelle & Mamadou Daffé* Continuous culture of Mycobacteria. Joanna Bacon*, Kim Hatch Measuring Minimum Inhibitory Concentrations in Mycobacteria. Frederick A Sirgel, Ian JF Wiid and David Paul van Helden* Rapid screening of inhibitors of Mycobacterium tuberculosis growth using tetrazolium salts. Anita G. Amin, Shiva K. Angala, Delphi Chatterjee and Dean C. Crick* Electroporation of Mycobacteria. Renan Goude* and Tanya Parish Ins and outs of mycobacterial plasmids. Farahnaz Movahedzadeh* and Wilbert Bitter The use of Temperature Sensitive Plasmids in Mycobacteria. Damien Portevin, Wladimir Malaga and ChristopheGuilhot* Heterologous Expression of Genes in Mycobacteria. James A. Triccas* and Anthony A. Ryan Inducible expression systems for mycobacteria. Christopher M. Sassetti* Assaying promoter activity using LacZ and GFP as reporters. Paul Carroll* and Jade James Construction of unmarked deletion mutants in mycobacteria. Houhui Song, Frank Wolschendorf and Michael Niederweis* Construction of targeted mycobacterial mutants by homologous recombination. Sharon L. Kendall* and Rosangela Frita Phage Transposon Mutagenesis. M. Sloan Siegrist and Eric J. Rubin* Gene Essentiality Testing in Mycobacterium smegmatis using Specialized Transduction. Apoorva Bhatt* and William R. Jacobs Jr. Gene switching and essentiality testing. Amanda Claire Brown* Insertion element IS6110-based Restriction Fragment Length Polymorphism (RFLP) Genotyping of Mycobacterium tuberculosis. Robin M. Warren*, Paul D. van Helden, Nicolaas C. Gey van Pittius Typing M. tuberculosis using VNTR analysis. TJ Brown, VN Nikolayevskyy and FA Drobniewski. Molecular Detection of Drug Resistant Mycobacterium tuberculosis with a scanning-frame oligonucleotide microarray. Dmitriy V. Volokhov, Vladimir E. Chizhikov, Steven Denkin, Ying Zhang* MycoDB: an online database for comparative genomics of the Mycobacteria and related organisms. Roy R. Chaudhuri* Environmental amoeba and mycobacterial pathogenesis. Melanie Harriff and Luiz E. Bermudez*

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