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Methods Mol Biol. 2019;2016:17-27. doi: 10.1007/978-1-4939-9570-7_2.

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

Author information

1
Department of Microbiology, Oregon State University, Corvallis, OR, USA.
2
Department of Food Science and Technology, Oregon State University, Corvallis, OR, USA.
3
Department of Food Science, Center for Food Safety, University of Arkansas, Fayetteville, AR, USA.
4
Cell and Molecular Biology Program, University of Arkansas, Fayetteville, AR, USA.
5
Department of Animal Science, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, FL, USA.
6
Department of Food Science and Technology, Oregon State University, Corvallis, OR, USA. sihong.park@oregonstate.edu.

Abstract

Transposons are elements widely dispersed among organisms which are able to move and replicate fragments of genomes. The extensive variability in transposons present in most organisms requires extensive identification and interpretation of the resulting transposon mutants after transposon mutagenesis. However, much of this is reliant on utilizing randomness characteristics of transposon to identify essential genes for the organism of interest. Integration of the transposon mutant approach with commercialized next-generation sequencing (NGS) technology has helped to advance transposon identification by sequencing millions of reads generated from a single run on an NGS platform. Transposon sequencing is defined as a gene sequencing methodology that allows for the identification of nonessential genes and the determination of gene function using a random transposon insertional mutagenesis followed by massively parallel sequencing. The detailed protocol will be outlined in this chapter. The genomic DNA integrated with the transposons is sequenced using an NGS platform in order to determine the location of each mutation.

KEYWORDS:

Next-generation sequencing (NGS); Salmonella Heidelberg; Transposon mutant; Whole-genome sequencing (WGS)

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