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About this book
This manual teaches theoretical and practical molecular genetic approaches to bacterial pathogenicity. Chapters on concepts, technologies and applications are followed by 15 experiments with Salmonella and Vibrio, in which protocols and expected findings are demonstrated and strategies for similar approaches to other bacteria are discussed. This manual is aimed at microbiologists in research, industrial and public health laboratories.
Overview: approaching genetic analysis of diverse bacteria. Bacterial pathogenesis: general concepts in bacterial pathogenesis; paradigms of bacterial pathogenesis - Salmonella typhimurium, Vibrio cholerae. Practical aspects of microbial genetics: genetic nomenclature; microbiological procedures; culture media; antibiotics, antibiotic resistance, and positive resistance, and positive and negative selections; bacterial physiology. Genetic mapping: genetic mapping of chromosomal genes; generalized transduction; genetic mapping by generalized transduction; deletion mapping. Mutants and their analysis: mutagenesis; broad host range allelic exchange systems; suppression; genetic complementation; polarity. Transposons and fusions: uses of transposons in bacterial genetics; bacteriophage mu; operon and gene fusions; operon and gene fusions with lacZ; operon fusions with uidA (gus), gene fusions with phoA, in vivo expression technology. Gene regulation: regulation of virulence genes; challenge phage; genetic analysis of DNA-protein interactions. Recombinant DNA: plasmids and conjugation; transformation and electrotransformation; basic molecular biology techniques; electrophoresis of DNA; polymerase chain reaction. Experiments. Appendices: strains, phage, and plasmids; culture media and supplements; strain colelctions; genetic exchange and mapping; enzyme assays; recombinant DNA methods; plasmid and transposon restriction maps.