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High-throughput Parallel Sequencing to Measure Fitness of Leptospira interrogans Transposon Insertion Mutants During Golden Syrian Hamster Infection
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1Veterans Affairs Greater Los Angeles Healthcare System, 2Departments of Medicine,David Geffen School of Medicine at University of California Los Angeles, 3Departments of Urology,David Geffen School of Medicine at University of California Los Angeles, 4Departments of Microbiology, Immunology, and Molecular Genetics,University of California Los Angeles
Chapitres
- 00:05Titre
- 01:17Creation of the Transposon Mutant Library: Transfer of the Transposon into Leptospira spp. by Conjugation
- 03:56Animal Experiment
- 04:57Making the Genomic Library
- 09:24Results: Challenged Hamsters
- 10:59Conclusion
We describe here a technique that combines transposon mutagenesis with high-throughput sequencing to identify and quantify transposon leptospiral mutants in tissues after a challenge of hamsters. This protocol can be used to screen mutants for survival and dissemination in animals and can also be applied to in vitro studies.
