Figure 1

Growth kinetics of BCG MDP1 cKD (A) Position of hupB target sequence start site shown by purple vertical lines for MDP1_ #1 and MDP1 #2 cKD BCG. Numbering indicates first nucleotide on the target sequence relative to annotated start coding sequence. Arrow indicates position of transcription start site. BCG growth kinetics depicted by (B) optical density (OD₆₀₀ nm) and (C) CFU over 21 days. Bacteria were cultured in 7H9/ADC medium supplemented with 200ng/ml ATc every 48 hours to induce sgRNA expression. At indicated time points culture aliquots were harvested to determine the OD and enumerate CFU after culturing on 7H10/OADC agar. Data represent mean ± SE from three biological replicates. Statistical differences between VC ATc and MDP1-cKD BCG were assessed by unpaired Welch’s T test, *P<0.05 **P<0.01 ***P<0.001; ^#p<0.05 indicate differences between MDP1_#1-cKD and MDP1_#2-cKD. (D) Representative western blot images cropped from different gels delineated with black border lines confirming the expression of MDP1: InhA serves as a loading control. Full-length blot images in Supplementary Fig. 3. 1-VC ATc, 2-MDP1_#1-cKD, 3-MDP1_#2-cKD. Image is representative of three experiments.