Book chapter
Generation and Screening of an Insertion Sequencing-Compatible Mutant Library of Campylobacter jejuni
Campylobacter jejuni, pp.257-272
Methods in Molecular Biology, Humana Press Inc
01/01/2017
DOI: 10.1007/978-1-4939-6536-6_21
PMID: 27885613
Abstract
The advent of next-generation sequencing technology has enabled experimental approaches to characterize large, complex populations of DNA molecules with high resolution. Included among these are methods to assess populations of transposon insertion libraries for the fitness cost of any particular mutant allele after applying selection to a population. These approaches have proven invaluable for identifying genetic factors that influence survival of bacterial pathogens within different environments, including animal hosts. One such method, termed insertion-site sequencing (INSeq), was designed to generate a 16 bp fragment of transposon-flanking genomic DNA captured during the protocol, which then serves as the substrate for massively parallel sequencing. Here we describe the generation of a transposon mutant library of Campylobacter jejuni amenable to INSeq and its use in identifying colonization determinants in a day-of-hatch chicken colonization model.
Details
- Title: Subtitle
- Generation and Screening of an Insertion Sequencing-Compatible Mutant Library of Campylobacter jejuni
- Creators
- Jeremiah G. Johnson - Michigan State UniversityVictor J. DiRita - Michigan State University
- Contributors
- J Butcher (Editor)A Stintzi (Editor)
- Resource Type
- Book chapter
- Publication Details
- Campylobacter jejuni, pp.257-272
- Publisher
- Humana Press Inc; TOTOWA
- Series
- Methods in Molecular Biology
- DOI
- 10.1007/978-1-4939-6536-6_21
- PMID
- 27885613
- eISSN
- 1940-6029
- ISSN
- 1064-3745
- Number of pages
- 16
- Grant note
- R01 AI069383; T32 AI007528 / NIAID NIH HHS; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of Allergy & Infectious Diseases (NIAID) T32AI007528 / NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of Allergy & Infectious Diseases (NIAID)
- Language
- English
- Date published
- 01/01/2017
- Academic Unit
- Microbiology and Immunology
- Record Identifier
- 9984696583402771
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