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KMID : 0880220210590121067
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Journal of Microbiology
2021 Volume.59 No. 12 p.1067 ~ p.1074
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Transposon insertion site sequencing (TIS) of Pseudomonas aeruginosa
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Cho Hong-Baek
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Abstract
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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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KEYWORD
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transposon insertion site sequencing, transposon mutagenesis, sequencing library, negative screening, fitness, synthetic lethality
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