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This book addresses cutting-edge techniques for researching transposon mutagenesis, an approach for identifying individual gene contributions to the phenotypic characteristics of a particular microorganism. The volume begins with methods for specific microorganisms and include protocols for individual microorganisms ranging from pathogens such as Salmonella to Bifidobacterium, a microorganism considered beneficial to humans and animals. The final section addresses more general protocols including plasmid transfer and bioinformatic tools as well as novel applications of transposon methodologies such as transposon-aided capture of antibiotic resistant plasmids. 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. 
Authoritative and practical, Microbial Transposon Mutagenesis: Protocols and Applications serves as a valuable reference for scientists seeking to apply transposon mutagenesis to microbial genetic analyses and functionality.

Microbial Transposon Mutagenesis: Protocols and Applications

Part I: Transposon Protocols for Specific Microorganisms

 

1. Preparation of Transposon Library and Tn-Seq Amplicon Library for Salmonella Typhimurium

            Sardar Karash, Tieshan Jiang, Deepti Samarth, Reena Chandrashekar, and

Young Min Kwon

 

2. Application of Whole Genome Sequencing to Transposon Mutants of Salmonella Heidelberg

            Bryna Rackerby, Sang In Lee, Ian Moppert, Steven C. Ricke, KwangCheol C. Jeong, and Si Hong Park

 

3. Construction of a Sequence-Defined Transposon Mutant Library in Staphylococcus aureus 

            Jennifer L. Endres, Vijaya Kumar Yajjala, Paul D. Fey, and Kenneth W. Bayles

 

4. Transposon Mutagenesis in Streptococcus Species

            Martin Nilsson, Michael Givskov, and Tim Tolker-Nielsen

 

5. Implementation of Transposon Mutagenesis in Bifidobacterium

            Lorena Ruiz and Douwe van Sinderen

 

6. Transposon Mutagenesis of Listeria monocytogenes

            Oindrila Paul, Damayanti Chakravarty, and Janet R. Donaldson

 

7. Transposon Mutagenesis of Foodborne Pathogenic Esherichia coli

            Supraja Puttamreddy and F. Chris Minion

 

8. Methods for Transposon Mutagenesis in Proteus mirabilis

            Philip N. Rather

 

9. Mutagenesis of Vibrio fischeri and Other Marine Bacteria Using Hyperactive Mini-Tn5 Derivatives

            Julie L. Stoudenmire, Michael Black, Pat M. Fidopiastis, and Eric V. Stabb

 

10. Transposon Mutagenesis of Bacteroides fragilis

            Yaligara Veeranagouda, Fasahath Husain, and Hannah M. Wexler

 

11. Transposon Mutagenesis in Mycobacterium avium Subspecies Paratuberculosis

            John P. Bannantine, Denise K. Zinniel, and Raúl G. Barletta

 

Part II: Transposon General Protocols and Applications

 

12. Protocols of Conjugative Plasmid Transfer in Salmonella: Plate, Broth, and Filter Mating Approaches

            Bijay K. Khajanchi, Pravin R. Kaldhone, and Steven L. Foley

 

13. Construction of DNA Barcode-Tagged Salmonella Strains

            Yichao Yang, Reena Chandrashekar, Steven C. Ricke, and Young Min Kwon

 

14. Transposon-Aided Capture of Antibiotic Resistance Plasmids from Complex Samples

            Sarah Delaney, Richard Murphy, and Fiona Walsh

 

15. Efficient Gene Deletion Method for Listeria monocytogenes

            Hossam Abdelhamed, Attila Karsi, and Mark L. Lawrence

 

16. Whole Genome Identification and Characterization of Bacterial Insertion Sequences Using Bioinformatic Tools

            Kody A. Bassett, Melanie R. Mormile, and Ronald L. Frank