Table 1.
Plasmids and M. tuberculosis strains used in this work
| Plasmid or strain | Antibiotic marker or resistance | Description or genotype | Mycolic acid phenotype |
|---|---|---|---|
| Plasmids | |||
| pMV306kan | Kanamycin | attB-integrating empty vector | |
| pDB22 | Zeocin | attB-integrating vector expressing mmaA1, mmaA2, mmaA3, and mmaA4 | |
| pDB32 | Kanamycin | Multicopy, episomal empty expression vector | |
| pDB46 | Kanamycin | Multicopy, episomal expression vector, based on pDB32, expressing mmaA1, mmaA2, mmaA3, mmaA4, pcaA, and cmaA2 | |
| pDB49 | Kanamycin | attB-integrating vector expressing mmaA3 and mmaA4 | |
| pDB60 | Streptomycin | attB-integrating empty vector | |
| pDB61 | Streptomycin | attB-integrating vector expressing mmaA3 and mmaA4 | |
| pDB90 | Kanamycin | attB-integrating vector, based on pMV306kan, expressing mmaA1, mmaA2, mmaA3, mmaA4, pcaA, and cmaA2 | |
| pDB111 | Kanamycin | attB site integration vector expressing mmaA3 and mmaA4; does not carry an integrase gene; can be inserted into bacteria only with another integrase-containing plasmid at the attB site; cannot be lost spontaneously | |
| Strains | |||
| Erdman | None | Wild type | Wild type |
| MGM1953 | Hygromycin, Zeocin | ΔmmaA1-4::loxP-hyg-loxP attB::mmaA1-4 (zeocin) | Wild type |
| MGM1960 | Hygromycin, Zeocin | ΔmmaA1-4::loxP ΔpcaA/umaA1::hyg attB::mmaA1-4 | Fully defective in proximal alpha cyclopropanation |
| MGM1961 | Hygromycin, streptomycin | ΔmmaA1-4::loxP ΔpcaA/umaA1::hyg attB::mmaA3-4 (strep) | Fully defective in proximal alpha cyclopropanation, partial defect in distal alpha cyclopropanation, fully defective in trans-cyclopropanation of methoxy- and ketomycolates |
| MGM1962 | Hygromycin, streptomycin | ΔmmaA1-4::loxP ΔpcaA/umaA1::hyg attB::(strep) | No oxygenated mycolates; alpha mycolates are fully defective for proximal cyclopropanation and partially defective for distal cyclopropanation |
| MGM1985 | Streptomycin | attB::(strep) | Wild type |
| MGM1990 | Hygromycin, Zeocin, streptomycin | ΔmmaA1-4::loxP attB::mmaA3-4 (strep) ΔpcaA/umaA1::hyg ΔcmaA2::zeo | Has all 3 classes (alpha, methoxy, keto) but no cyclopropane rings |
| MGM1991 | Hygromycin, Zeocin, streptomycin | ΔmmaA1-4::loxP attB::(strep) ΔpcaA/umaA1::hyg ΔcmaA2::zeo | Has no oxygenated mycolates, only alpha mycolates; has no cyclopropane rings |
| MGM1995 | Hygromycin, Zeocin | ΔmmaA1-4::loxP ΔpcaA/umaA1::hyg ΔcmaA2::zeo | Identical to MGM1991 |
| MGM1998 | Hygromycin, Zeocin, kanamycin | ΔmmaA1-4::loxP attB::mmaA3-4 (kan) ΔpcaA/umaA1::hyg ΔcmaA2::zeo | Identical to MGM1990 but no integrase at attB |