TL;DR
A transposon-sequencing protocol (Tn-Seq) that combines random transposon mutagenesis with high-throughput sequencing to simultaneously assess fitness changes across a large collection of L. interrogans mutants under defined conditions is described.
Tn-seq is particularly useful as a high-throughput method to assign function to function-unknown genes, which have increased steadily with the abundance of newly sequenced bacterial genomes.
Together, this strategy provides a powerful framework to uncover the genes essential for fungal development and virulence within the host.
ABSTRACT Although the original Hermes transposon mutagenesis system has proven successful for saturated insertional mutagenesis in Candida glabrata, its reliance on uracil auxotrophy may limit its applicability under nutrient-restricted host environments. To overcome this limitation, Zaeske et al. (Microbiol Spectr 14:...
The findings may inform the identification of potential therapeutic targets for multidrug-resistant K. pneumoniae and the identification of genes essential for growth on laboratory media.
Bacteriophage genomes contain large fractions of uncharacterized genes, motivating high-throughput functional screens to illuminate their roles in host interaction and cytotoxicity. Here, we performed a genome-wide overexpression assay for a Pseudomonas putida KT2440-infecting phage, Emajogi, and identified eight cytot...
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