Figure 1
Growth inhibition by VapC-mt11. (A) M. smegmatis mc2155 harboring the anhydrotetracycline (ATc) inducible vector pMC1s with VapC-mt11 was grown at 37 °C in 7H9 + TW80 + AND supplemented with 25 ug/mL kanamycin. Cultures were split and one culture was induced by the addition of 200 ng/mL ATc. Time points for uninduced (♦) and induced (
) samples were collected up to 6 h. Growth curve shown is representative of the trend documented in three independent experiments. (B) M. tuberculosis H37Rv transformed with pMC1s-VapCmt11 were grown at 37 °C in 7H9-TW80-ADN medium containing 30% spent medium and supplemented with 25 ug/mL kanamycin. Cultures were separated into uninduced (▲) and induced (
). Induced cultures were supplemented with 200 ng/ml ATc and additional ATc was added every 48 h to maintain the ATc concentration between 100 and 250 ng/ml. Data points represent the average of three independent experiments; error bars represent the S.D. (C) Plate efficiency assay for M. tuberculosis H37Rv cells harboring pMC1s-VapC-mt11 between uninduced (gray) and induced (red). Transformants were plated on 7H9 + TW80 + AND agar with 25 ug/mL kanamycin with or without 500 ng/mL of ATc and incubated for 3 weeks at 37 °C. Error bars represent the S.D. for biological and technical replicates.
