Mycobacteria Protocols

by Parish, Tanya; Brown, Amanda Claire

• ISBN13: 9781588298898
• ISBN10: 1588298892
• Edition: 2nd
• Format: Hardcover
• Copyright: 2008-11-30
• Publisher: Humana Pr Inc

Summary

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. Book jacket.

Table of Contents

Preface	p. v
Contributors	p. xiii
Isolation of Mycobacterium Species Genomic DNA	p. 1
Isolation of Mycobacterial RNA	p. 13
Lipoglycans of Mycobacterium tuberculosis: Isolation, Purification, and Characterization	p. 23
Analyzing Lipid Metabolism: Activation and [beta]-Oxidation of Fatty Acids	p. 47
Analysis of Lipid Biosynthesis and Location	p. 61
Whole Genome Analysis Using Microarrays	p. 83
Use of DNA Microarrays to Study Global Patterns of Gene Expression	p. 95
Two-Dimensional Gel Electrophoresis-Based Proteomics of Mycobacteria	p. 111
Transport Assays and Permeability in Pathogenic Mycobacteria	p. 143
Continuous Culture of Mycobacteria	p. 153
Measuring Minimum Inhibitory Concentrations in Mycobacteria	p. 173
Rapid Screening of Inhibitors of Mycobacterium tuberculosis Growth Using Tetrazolijm Salts	p. 187
Electroporation of Mycobacteria	p. 203
Ins and Outs of Mycobacterial Plasmids	p. 217
The Use of Temperature-Sensitive Plasmids in Mycobacteria	p. 229
Heterologous Expression of Genes in Mycobacteria	p. 243
Inducible Expression Systems for Mycobacteria	p. 255
Assaying Promoter Activity Using LacZ and GFP as Reporters	p. 265
Construction of Unmarked Deletion Mutants in Mycobacteria	p. 279
Construction of Targeted Mycobacterial Mutants by Homologous Recombination	p. 297
Phage Transposon Mutagenesis	p. 311
Gene Essentiality Testing in Mycobacterium smegmatis Using Specialized Transduction	p. 325
Gene Switching and Essentiality Testing	p. 337
Insertion Element IS6110-Based Restriction Fragment Length Polymorphism Genotyping of Mycobacterium tuberculosis	p. 353
Typing Mycobacterium tuberculosis Using Variable Number Tandem Repeat Analysis	p. 371
Molecular Detection of Drug-Resistant Mycobacterium tuberculosis with a Scanning-Frame Oligonucleotide Microarray	p. 395
MycoDB: An Online Database for Comparative Genomics of the Mycobacteria and Related Organisms	p. 419
Environmental Amoebae and Mycobacterial Pathogenesis	p. 433
Index	p. 443
Table of Contents provided by Ingram. All Rights Reserved.