Transposon insertion site sequencing (TIS) of Pseudomonas aeruginosa

Hongbaek Cho

doi:10.1007/s12275-021-1565-y

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Transposon insertion site sequencing (TIS) of Pseudomonas aeruginosa: Hongbaek Cho; Journal of Microbiology 2021;59(12):1067-1074.
DOI: https://doi.org/10.1007/s12275-021-1565-y
Published online: December 4, 2021

Department of Biological Sciences, College of Natural Sciences, Sungkyunkwan University, Suwon 16419, Republic of Korea

Corresponding author: Hongbaek Cho , Tel: +82-31-299-4498,

Received: 25 October 2021 • Revised: 11 November 2021 • Accepted: 12 November 2021

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Abstract

Transposon insertion site sequencing (TIS) is a technique that determines the insertion profile of a transposon mutant library by massive parallel sequencing of transposon-genomic DNA junctions. Because the transposon insertion profile reflects the abundance of each mutant in the library, it provides information to assess the fitness contribution of each genetic locus of a bacterial genome in a specific growth condition or strain background. Although introduced only about a dozen years ago, TIS has become an important tool in bacterial genetics that provides clues to study biological functions and regulatory mechanisms. Here, I describe a protocol for generating high density transposon insertion mutant libraries and preparing Illumina sequencing samples for mapping the transposon junctions of the transposon mutant libraries using Pseudomonas aeruginosa as an example.

Keywords: transposon insertion site sequencing, transposon mutagenesis, sequencing library, negative screening, fitness, synthetic lethality

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Transposon insertion site sequencing (TIS) of Pseudomonas aeruginosa

J. Microbiol. 2021;59(12):1067-1074. Published online December 4, 2021

DOI: https://doi.org/10.1007/s12275-021-1565-y

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