Mycobacteria Protocols

Methods to study the biology of mycobacteria are still very much in need. This new 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.

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™ 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 Includes supplementary material: sn.pub/extras

Inhalt
Isolation of Mycobacterium Species Genomic DNA.- Isolation of Mycobacterial RNA.- Lipoglycans of Mycobacterium tuberculosis: Isolation, Purification, and Characterization.- Analyzing Lipid Metabolism: Activation and ?-Oxidation of Fatty Acids.- Analysis of Lipid Biosynthesis and Location.- Whole Genome Analysis Using Microarrays.- Use of DNA Microarrays to Study Global Patterns of Gene Expression.- Two-Dimensional Gel Electrophoresis-Based Proteomics of Mycobacteria.- Transport Assays and Permeability in Pathogenic Mycobacteria.- Continuous Culture of Mycobacteria.- Measuring Minimum Inhibitory Concentrations in Mycobacteria.- Rapid Screening of Inhibitors of Mycobacterium tuberculosis Growth Using Tetrazolium Salts.- Electroporation of Mycobacteria.- Ins and Outs of Mycobacterial Plasmids.- The Use of Temperature-Sensitive Plasmids in Mycobacteria.- Heterologous Expression of Genes in Mycobacteria.- Inducible Expression Systems for Mycobacteria.- Assaying Promoter Activity Using LacZ and GFP as Reporters.- Construction of Unmarked Deletion Mutants in Mycobacteria.- Construction of Targeted Mycobacterial Mutants by Homologous Recombination.- Phage Transposon Mutagenesis.- Gene Essentiality Testing in Mycobacterium smegmatis Using Specialized Transduction.- Gene Switching and Essentiality Testing.- Insertion Element IS6110-Based Restriction Fragment Length Polymorphism Genotyping of Mycobacterium tuberculosis.- Typing Mycobacterium tuberculosis Using Variable Number Tandem Repeat Analysis.- Molecular Detection of Drug-Resistant Mycobacterium tuberculosis with a Scanning-Frame Oligonucleotide Microarray.- MycoDB: An Online Database for Comparative Genomics of the Mycobacteria and Related Organisms.- Environmental Amoebae and Mycobacterial Pathogenesis.