The katG gene was knocked out by introducing a premature stop codon at Gln3 (CAA to TAA) using the CTBEevolve plasmid. The strains were grown in Middlebrook 7H9 broth with OADC to the stationary phase. A fraction of the cells was plated onto the Middlebrook 7H10 agar plate supplemented with OADC without INH and another fraction was plated onto a Middlebrook 7H10 agar supplemented with OADC and INH (16 μg/mL). The plates were sealed by parafilm and incubated for 20 days at 37°C.