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Immunology and Infection

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JoVE Journal Immunology and Infection
High-throughput Parallel Sequencing to Measure Fitness of Leptospira interrogans Transposon Insertion Mutants During Golden Syrian Hamster Infection
 

High-throughput Parallel Sequencing to Measure Fitness of Leptospira interrogans Transposon Insertion Mutants During Golden Syrian Hamster Infection

Article DOI: 10.3791/56442-v • 11:50 min • December 18th, 2017
December 18th, 2017 •
1,2, 1,2, 1,2, 1,2,3,4
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

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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.

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High-throughput Parallel Sequencing Fitness Measurement Leptospira Interrogans Transposon Insertion Mutants Golden Syrian Hamster Infection In Vivo Fitness Defects Virulence Genes Colonization Dissemination Survival Large Number Of Mutants Limited Number Of Animals Acute Leptospirosis Chronic Infection Models In Vitro Studies Filtration Unit Leptospira Culture E.coli Culture EMJH Plate DAP
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