| Title | Transposon insertion site sequencing (TIS) of Pseudomonas aeruginosa |
|---|---|
| Author | Hongbaek Cho* |
| Address | Department of Biological Sciences, College of Natural Sciences, Sungkyunkwan University, Suwon 16419, Republic of Korea |
| Bibliography | Journal of Microbiology, 59(12),1067–1074, 2021, |
| DOI | 10.1007/s12275-021-1565-y |
| Key Words | transposon insertion site sequencing, transposon mutagenesis, sequencing library, negative screening, fitness, synthetic lethality |
| 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. |