FIG 2 .

Competitive infection against the wild-type strain confirms the fitness defect of mutants selected by InSeq. Insertion-deletion mutants of six genes selected and one gene not selected by CEDER analysis (VK055_468) of InSeq fitness were constructed using Lambda Red recombinase and used to coinfect mouse lungs with the wild type (WT), or WT carrying pACYC184 where noted, at a 1:1 ratio in a total inoculum of 1 × 10⁶ CFU. Mean competitive index (log₁₀) for each mutant compared to the wild type at 24 h is shown. *, P < 0.05; **, P < 0.01; ***, P < 0.001, by one-sample t test on log₁₀-transformed data (≥4 mice per group); ###, P < 0.001 by two-sample t test.