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*
Due to the rising threat of tuberculosis and other mycobacterial infections, methods to study the biology of the mycobacteria and to improve diagnostic, therapeutic and preventative reagents are still very much in need. Mycobacteria Protocols, Second Edition updates and refines the methods of the well-received first edition while adding newly developed methods culled from the most cutting-edge research in the field. Ranging from the basics of sub-cellular fractionation to advanced methods using specialized growth conditions and whole genome, transcriptome and proteome analysis, the chapters conform to the Methods in Molecular Biology(TM) series format, providing step-by-step laboratory protocols, lists of necessary materials and reagents, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and up-to-date, Mycobacteria Protocols, Second Edition is the perfect resource to promote and stimulate further research into these intriguing, important and most fractious of bacteria.
Fully updated from the first edition to offer cutting edge methods for manipulation of Mycobacteria
Includes methods for preparation and analysis of sub-cellular fractions
Provides description of genetic systems and instructions for manipulating the genome
Contains techniques for studying gene expression, singly and globally, and updated techniques for mutagenesis and gene essentiality testing
With detailed protocols for genomic, transcriptomic and proteomic analysis, and new molecular methods for typing and drug susceptibility profiling