Figure 5. MSMEG_6382 is essential for growth of M. smegmatis in LB broth.

6382CKO was cultured at 30°C in LB containing Kn and Sm. At saturation, 5 ml was used to inoculate 200 ml of LB/Kn medium that had been prewarmed at the permissive (30°C •) or non-permissive (42°C ▪) temperature. Incubation was continued at the two temperatures and both cultures were sampled regularly with serial dilutions plated on LB plates containing Kn to determine colony forming units (CFUs) per ml. A wild-type M. smegmatis mc²155 strain containing the kanamycin resistance plasmid pMV261 was included as a control (30°C ▴, 42°C ▾). Unbroken lines represent the 6382CKO strain while broken lines represent the wild type (pMV261) control strain. These data represent means of triplicate samples ± standard deviation and are representative of three independent experiments.