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-receivedfirst 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 inMolecular Biology™ series format, providing step-by-step laboratory protocols, lists of necessary materials and reagents, and tips on troubleshooting and avoiding known pitfalls. 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. INDICE: From the contents Isolation of Mycobacterium ssp. Genomic DNA.- Isolation of Mycobacterial RNA.- Lipoglycans of Mycobacterium tuberculosis: isolation, purification and characterization.- Analysing lipid metabolism: activation and ß-oxidation of fatty acids.- Analysis of lipid biosynthesis and location.- Whole genome analysis using microarrays.- Use of DNA microarrays to studyglobal patterns of gene expression.- Two-dimensional gel electrophoresis-based proteomics of mycobacteria.- Transport assays and permeability in pathogenicmycobacteria.- 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.
- ISBN: 978-1-58829-889-8
- Editorial: Humana
- Encuadernacion: Cartoné
- Páginas: 490
- Fecha Publicación: 01/07/2008
- Nº Volúmenes: 1
- Idioma: Inglés