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Transposon insertion site sequencing (TIS) of Pseudomonas aeruginosa

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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.

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Acknowledgements

This work was supported by the National Research Foundation of Korea (NRF) Grant (NRF- 2019R1A2C1002648)

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

    Hongbaek Cho

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  1. Hongbaek Cho
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Cho, H. Transposon insertion site sequencing (TIS) of Pseudomonas aeruginosa. J Microbiol. 59, 1067–1074 (2021). https://doi.org/10.1007/s12275-021-1565-y

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