Transposon Insertion Sequencing to study the genetic interactions between V. cholerae WT cells and two mutant strains in absence or presence of subinhibitory doses of tobramycin

This study was conducted to study which genes or functions become important or expendable in two V. cholerae mutants compared to the WT. This was assessed directly by conjugating a Mariner Transposon with WT or mutant cells and obtaining a bank of hundreds of thousands of mutants which we consider as the T=0. We then subjected these banks to 16 generations in rich media with or without 50% MIC of tobramycin (which we call T=16gen). At both T0 and T16gen we extracted the genomic DNA of these banks and sequenced it to identify the regions where the transposon is inserted in each strain at the different conditions. For each strain, three biological replicates were used. J085 strain corresponds to V. cholerae WT strain, H505 corresponds to the vchM (vca0198) mutant and M092 corresponds to the double mutant (vca0762-vca0763).