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This detailed book addresses the continual development and application of new methods and protocols to understand the biology of the pathogen Neisseria gonorrhoeae. Beginning with gonococcus biology and antimicrobial resistance, the volume continues with areas of study such as producing major gonococcal antigens to high purity, genetic transformation of gonococci and strategies for global RNA sequencing, evaluation of potential vaccine/drug targets and the assessment of hypothetical protein function, as well as a number of methods and protocols for studying the biological interactions of the gonococcus and the host. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and cutting-edge, Neisseria gonorrhoeae: Methods and Protocols provides researchers with a foundation for progress toward the eventual control of this disease and a significant reduction in global case numbers of gonorrhea.
List of contents
1. Biology of the Gonococcus: Disease and Pathogenesis.- 2. How to identify Exposed Women who are infected with Neisseria gonorhoeae.- 3. Antimicrobial resistance in Neisseria gonorrhoeae and treatment of gonorrhoea.- 4. Prediction of minimum inhibitory concentrations of antimicrobials for Neisseria gonorrhoeae using whole-genome sequencing.- 5. Gonococcal Colony Typing.- 6. Preparation of lipooligosaccharide (LOS) from Neisseria gonorrhoeae.- 7. Purification of Type IV Pili and Pilin Subunits.- 8. Peptidoglycan Composition in Neisseria.- 9. The isolation of naturally released gonococcal outer membrane vesicles as vaccine antigens.- 10. Transformation in Neisseria gonorrhoeae.- 11. Strategies for Global RNA Sequencing of the Human Pathogen Neisseria gonorrhoeae.- 12. Bioinformatics Workflow for Gonococcal Proteomics.- 13. Phenotypic MicroArray screening of Neisseria gonorrhoeae in chemically defined liquid medium.- 14. Generation of metal-depleted conditions for in vitro growth of Neisseria gonorrhoeae.- 15. Cloning, expression and purification of recombinant Neisseria gonorrhoeae proteins.- 16. Complement-dependent serum bactericidal assays for Neisseria gonorrhoeae.- 17. Basic methods for examining Neisseria gonorrhoeae interactions with host cells in vitro.- 18. Use of Human Monocyte-Derived Macrophages to Study Neisseria gonorrhoeae Infection.- 19. Protocols to interrogate the interactions between Neisseria gonorrhoeae and primary human neutrophils.- 20. Analysis of Host Responses to Neisseria gonorrhoeae using a Human Three-Dimensional Endometrial Epithelial Cell Model.- 21. In vitro models of eye infection with Neisseria gonorrhoeae.- 22. Use of Human Fallopian Tube Organ in Culture (FTOC) and Primary Fallopian Tube Epithelial Cells (FTEC) to Study the Biology of Neisseria gonorrhoeae infection.- 23. A Natural Mouse Model for Neisseria Persistent Colonization.- 24. Female Mouse Model of Neisseria gonorrhoeae Infection.- 25. Experimental Human Infection with Neisseria gonorrhoeae.
Summary
This detailed book addresses the continual development and application of new methods and protocols to understand the biology of the pathogen Neisseria gonorrhoeae. Beginning with gonococcus biology and antimicrobial resistance, the volume continues with areas of study such as producing major gonococcal antigens to high purity, genetic transformation of gonococci and strategies for global RNA sequencing, evaluation of potential vaccine/drug targets and the assessment of hypothetical protein function, as well as a number of methods and protocols for studying the biological interactions of the gonococcus and the host. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and cutting-edge, Neisseria gonorrhoeae: Methods and Protocols provides researchers with a foundation for progress toward the eventual control of this disease and a significant reduction in global case numbers of gonorrhea.
