Pseudomonas Methods and Protocols

In Pseudomonas aeruginosa, expert researchers in the field detail many of the methods which are now commonly used to study this fascinating microorganism. Chapters include microbiological methods to high-throughput molecular techniques that have been developed over the last decade. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols and key tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Pseudomonas aeruginosa aids in the continuing study of new and cutting edge findings.

Gene Transfer: Transduction.- Gene Transfer: Transformation/Electroporation.- Gene transfer: Conjugation.- Pseudomonas Bacteriophage Isolation and Production.- Genotyping Methods.- Drug Susceptibility Testing by Dilution Methods.- Plate-based Assay for Swimming Motility in Pseudomonas aeruginosa.- Plate-based Assay for Swarming Motility in Pseudomonas aeruginosa.- Motility Assay: Twitching Motility.- Qualitative and Quantitative Assays for Flagellum-mediated Chemotaxis.- Microscopic Analysis: Morphotypes and Cellular Appendages.- Determination of Lipolytic Activities.- Elastinolytic and proteolytic enzymes.- In vitro Assays to Monitor the Activity of Pseudomonas aeruginosa Type III Secreted Proteins.- Cell Fractionation.- Characterization of Molecular Interactions using Isothermal Titration Calorimetry.- Proteomic Analysis.- Membrane Proteomics of Pseudomonas aeruginosa.- Construction of Pseudomonas Two-hybrid Libraries for High-throughput Assays.- Biosensors for Qualitative and Semi-quantitative Analysis of Quorum Sensing Signal Molecules.- LC-MS/MS Quantitative Analysis of Quorum Sensing Signal Molecules.- LC/MS/MS-Based Quantitative Assay for the Secondary Messenger Molecule, c-di-GMP.- Metabolic Footprinting: Extracellular Metabolomic Analysis.- Pyoverdine and Pyochelin Measurements.- Measurement of Phenazines in Bacterial Cultures.- Extraction and Measurement of NAD(P)⁺ and NAD(P)H.- Cyanide Measurements in Bacterial Culture and Sputum.- Monitoring Iron Uptake by Siderophores.- Exopolysaccharide Quantification.- Liquid Chromatography/Mass Spectrometry for the Identification and Quantification of Rhamnolipids.- LPS quantitation Procedures.- Monitoring Lectin Interaction with Carbohydrates.- Mining the Pseudomonas Genome.- Identification of Bacterial Small RNAs by RNA Sequencing.- Gene Amplification and qRT-PCR.- The Standard European Vector architecture (SEVA) plasmid toolkit.- Chromosomal integration of transcriptional fusions.- A Method toCapture Large DNA Fragments from Genomic DNA.- Transposon Mutagenesis.- Site-directed Mutagenesis and Gene Deletion using Reverse Genetics.- Signature Tagged Mutagenesis.- Construction of a Pseudomonas aeruginosa Genomic Bank.- Strategy for Genome Sequencing Analysis and Assembly for Comparative Genomics of Pseudomonas Genomes.- Promoter Fusions with Optical Outputs in Individual Cells and in Populations.- Chromatin Immunoprecipitation for ChIP-chip and ChIP-Seq.- Transcriptional Analysis of Pseudomonas aeruginosa Infected Caenorhabditis elegans.- Methods for Studying Biofilm Formation: Flow Cells and Confocal Laser Scanning Microscop.- Biofilm Formation in the 96-well Microtiter Plate.- Methods for Studying Biofilm Dispersal in Pseudomonas aeruginosa.- Pseudomonas aeruginosa PA14 Pathogenesis in Caenorhabditis Elegans.- Assessing Pseudomonas Virulence using a Non-mammalian Host Model: Dictyostelium Discoideum.- Assessing Pseudomonas virulence with non-mammalian host: Galleria mellonella.- Assessing Pseudomonas Virulence with the Non-mammalian Host: Arabidopsis Thaliana.- Assessing Pseudomonas aeruginosa Persister/Antibiotic Tolerant Cells.- Assessing Pseudomonas Virulence with Non-mammalian Host: Zebrafish.- Assessing Pseudomonas Virulence with a Non-mammalian Host: Drosophila Melanogaster.- Assessing Pseudomonas Virulence using Host Cells.- Assessing Pseudomonas aeruginosa Virulence and the Host Response using Murine Models of Acute and Chronic Lung Infection.- Assessing Pseudomonas Virulence using Mammalian Models: Acute Infection Model.- Burn Mouse Models.