This volume of Methods in Enzymology presents methods for the isolation and identification of bacterial pathogens and associated virulence determinants. It will enable the researcher to determine if closed genes or isolated gene products are involved in virulence.

Table of Contents:
S.A. Morse and J.E. McDade, Recommendations for Working with Pathogenic Bacteria. Evaluations of Virulence in Animal Models: S. Welkos and A. O'Brien, Determination of Median Lethal and Infectious Doses in Animal Model Systems. D.J. Kopecko, Experimental Keratoconjunctivitis (Sereny) Assay. R.D. Shahin and J.L. Cowell, Mouse Respiratory Infection Models for Pertussis. B.A. Green, W.J. Doyle, and J.L. Cowell, Chinchilla Model of Experimental Otitis Media for Study of Nontypable Haemophilus influenzae Vaccine Efficacy. R.G. Rank and J.A. Whittum-Hudson, Animal Models for Ocular Infections. R.G. Rank, Animal Models for Urogenital Infections. M.G. Tauber and A. Zwahlen, Animal Models for Meningitis. T.E. Madden and J.G. Caton, Animal Models for Periodontal Disease. C.A. Genco and R.J. Arko, Animal Chamber Models for Study of Host-Parasite Interactions. D.F. Keren and M.A. Suckow, Animal Models for Immunoglobulin A Secretion. Species and Strain Identification: C.E. Frasch, Serogroup and Serotype Classification of Bacterial Pathogens. K. Nelson and R.K. Selander, Analysis of Genetic Variation by Polymerase Chain Reaction-Based Nucleotide Sequencing. R. Rudner, B. Studamire, and E.D. Jarvis, Determinations of Restriction Fragment Length Polymorphism in Bacteria Using Ribosomal RNA Genes. D. van Soolingen, P.E.W. de Haas, P.W.M. Hermans, and J.D.A. van Embden, DNA Fingerprintingof Mycobacterium tuberculosis. T.M. Schmidt and D.A. Relman, Phylogenetic Identification of Uncultured Pathogens Using Ribosomal RNA Sequences. Bacterial Cell Fractionation: H. Nikaido, Isolation of Outer Membranes. G. F.-L. Ames,Isolation and Purification of Periplasmic Binding Proteins. M.A. Apicella, J.M. Griffis, and H. Schneider, Isolation and Characterization of Lipopolysaccharides, Lipooligosaccharides, and Lipid A. R.S. Rosenthal and R. Dziarski, Isolation of Peptidoglycan and Soluble Peptidoglycan Fragments. V.A. Fischetti, Purification of treptococcal M Protein. T.B. May and A.M. Chakrabarty, Isolation and Assay of Pseudomonas aeruginosa Alginate. W.F. Vann and S.J. Freese, Purification of Escherichia coli K Antigens. Microbial Acquisition of Iron: C.D. Cox, Deferration of Laboratory Media and Assays for Ferric and Ferrous Ions. S.M. Payne, Detection, Isolation, and Characterization of Siderophores. J.B. Neilands, Effects of Iron Deprivation on Outer Membrane Protein Expression. J.B. Neilands, Identification and Isolation of Mutants Defective in Iron Acquisition. C.N. Cornelissen and P.F. Sparling, Identification of Receptor-Mediated Transferrin-Iron Uptake Mechanism in Neisseria gonorrhoeae. S.E.H. West, Isolation of Genes Involved in Iron Acquisition by Cloning and Complementation of Escherichia coli Mutants. Genetics and Regulation: J.F. Miller, Bacterial Transformation by Electroporation. V. de Lorenzo and K.N. Timmis, Analysis and Construction of Stable Phenotypes in Gram-Negative Bacteria with Tn5- and Tn10-Derived Minitransposons. R.A. Burne and R.G. Quivey, Jr., Use of Transposons to Dissect Pathogenic Strategies of Gram-Positive Bacteria. M.R. Kaufman and R.K. Taylor, Identification of Bacterial Cell-Surface Virulence Determinants with Tn pho A. A.M. Hooke, Temperature-Sensitive Mutants of Bacterial Pathogens: Isolation and Useto Determine Host Clearance and in Vivo Replication Rates. S. Stibitz, Use of Conditionally Counterselectable Suicide Vectors for Allelic Exchange. D.S. Toder, Gene Replacement in Psudomonas aeruginosa. P. Guerry, R. Yao, R.A. Alm, D.H. Burr, and T.J. Trust, Systems of Experimental Genetics for Campylobacter Species. J.M. Slauch, M.J. Mahan, and J.J. Mekalanos, In Vivo Expression Technology for Selection of Bacterial Genes Specifically Induced in Host Tissues. N.A.Zielinski, S. Roychoudhury, and A.M. Chakrabarty, Regulation of Alginate Gene Expression in Pseudomonas aeruginosa. D.W. Frank and D.G. Storey, Regulation of Expression of Pseudomonas Exotoxin A by Iron. C.L. Gardel and J.J. Mekalanos, Regulation of Cholera Toxin by Temperature, pH, and Osmolarity. M.S. Strom, D.N. Nunn, and S. Lory, Posttranslational Processing of Type IV Prepilin and Homologs by PilD of Pseudomonas aeruginosa. Enzyme and Toxin Assays: M.H. Mulks and R.J. Shoberg, Bacterial Immunoglobulin A1 Proteases. L. Rust, C.R. Messing, and B.H. Iglewski, Elastase Assays. M.S. Lantz and P. Ciborowski, Zymographic Techniques for Detection and Characterization of Microbial Proteases. J.D.Grubb, Assays for Bacterial Type I Collagenases. J.D. Grubb, Purification and Assays of Bacterial Gelatinases. W.L. Hynes and J.J. Ferretti, Assays for Hyaluronidase Activity. L. Passador and W. Iglewski, ADP-Ribosylating Toxins. S.F. Carroll and R.J. Collier, Photoaffinity Labeling of Active Site Residues in ADP-Ribosylating Toxins. J. Moss, S.-C. Tsai, and M. Vaughan, Activation of Cholera Toxin by ADP-Ribosylation Factors. T.G. Obrig, Toxins That Inhibit Host Protein Synthesis. G.E. Rowe and R.A. Welch, Assays of Hemolytic Toxins. A.L. Lobo and R.A. Welch, Identification and Assay of RTX Family of Cytolysins. M. Thelestam and I. Florin, Assay of Cytopathogenic Toxins in Cultured Cells. B.L. Kagan and Y. Sokolov, Use of Lipid Bilayer Membranes to Detect Pore Formation by Toxins. C.B. Saelinger and R.E. Morris, Uptake and Processing of Toxins by Mammalian Cells. Author Index. Subject Index.