FIGURE 3.

Extracellular growth and stress analysis results of recombinant M. smegmatis. (A) MS_Rv2650c and MS_Vec were inoculated into MB 7H9 broth and cultured. The OD₆₀₀ value was monitored at 4 h intervals for a total of 68 h. (B) Inoculate MS_Rv2650c and MS_Vec into 7H9 broth at pH 4.5 for 6 h at 37°C. The cultured solution of 0, 3, and 6 h was diluted and coated on MB 7H9 plate, and the number of single colonies was calculated after incubation at 37°C for 3–4 days. (C) MS_Rv2650c and MS_Vec were treated with 3% hydrogen peroxide at 37°C for 6 h in a cultured solution with an OD₆₀₀ about 0.5, and the cultured solution of 0, 3, and 6 h was diluted and coated on the MB 7H9 plate. The number of single colonies was calculated after incubation for 3–4 days at 37°C. (D) MS_Rv2650c and MS_Vec were treated with 2% SDS for 6 h at 37°C in a cultured solution with an OD₆₀₀ about 0.5, and the cultured solution of 0, 3, and 6 h was diluted and coated on the MB 7H9 plate. The number of single colonies was calculated after incubation for 3–4 days at 37°C. (E) MS_Rv2650c and MS_Vec were diluted and coated on 7H9 plates under the conditions of pH 4.5, 3% H₂O₂, 2% SDS, and cultured at 37°C for 3–4 days. Similar results were obtained in 3 independent experiments. (*p < 0.05; **p < 0.01; ***p < 0.001).