Browsing by Author "Block, Alisha M"

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    Tnseq screen of Mycobacterium tuberculosis Himar1 transposon library grown in MtbYM rich medium and treated with antitubercular drugs.
    (2022-08-15) Block, Alisha M; Tischler, Anna D; tischler@umn.edu; Tischler, Anna D; University of Minnesota Tischler Lab
    Transposon-sequencing (Tn-seq) is a powerful genetic tool that can identify changes in transposon (Tn) mutant frequency during a given test condition such as drug treatment during nutrient starvation. We created an arrayed library of Tn mutants in Mycobacterium tuberculosis Erdman, screened a subset of the library (~1,000 mutants) by Tn-seq, and identified 100+ Tn mutants with altered tolerance to antibiotics in starvation conditions. We individually retested three Tn mutants to confirm their Tn-seq phenotypes, rv0457c::Tn, ppgK::Tn and clpS::Tn, and all three mutants showed reproducible increased susceptibility to drug treatment during starvation. However, the phenotypes for two of Tn mutants, ppgK::Tn and clpS::Tn, were not complemented by the Tn-disrupted gene. Whole-genome sequencing revealed that both Tn mutants had single nucleotide polymorphisms that prevented PDIM production, which was responsible for the observed drug tolerance phenotypes. This repository contains all the Illumina sequencing data for the M. tuberculosis antibiotic tolerance Tn-seq experiments and whole-genome sequencing data for our lab WT Erdman strain and three Tn mutants identified in the Tn-seq screen that were individually retested.