Mtb H37Rv was used as virulent wild-type strain. BPL-DUC and TrxB2-DUC have been previously described (Tiwari et al., 2018 (link); Lin et al., 2016 (link)). BCG Pasteur was from ATCC strain TMC 1011 (#35734). Strains were cultured in liquid Middlebrook 7H9 medium supplemented with 0.2% glycerol, 0.05% Tween 80, and ADN (0.5% BSA, 0.2% dextrose, and 0.085% NaCl) and on Middlebrook 7H10 agar plates supplemented with 0.2% glycerol and Middlebrook OADC enrichment (Becton Dickinson). Antibiotics were added for selection of genetically modified strains at the following concentrations: hygromycin (50 µg/ml), kanamycin (25 µg/ml), and zeocin (25 µg/ml).
Protocol detail
Mycobacterial Strain Culture Conditions
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Corresponding Organization : Cornell University
Other organizations : Texas A&M University
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Variable analysis
independent variables
- Strain type (Mtb H37Rv, BPL-DUC, TrxB2-DUC, BCG Pasteur)
- Not explicitly mentioned
- Culture conditions (Middlebrook 7H9 medium supplemented with 0.2% glycerol, 0.05% Tween 80, and ADN; Middlebrook 7H10 agar plates supplemented with 0.2% glycerol and Middlebrook OADC enrichment)
- Antibiotics used for selection (hygromycin, kanamycin, zeocin)
- Positive control: Mtb H37Rv (virulent wild-type strain)
- Negative control: Not explicitly mentioned
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