TABLE 2.
Strains used in this study
| Strain | Relevant genotype or characteristics | Reference or constructiona |
|---|---|---|
| B. subtilis | ||
| PS832 | Wild type, Trp+ revertant of strain 168 | Laboratory stock |
| PS2061 | ΔprfA::spc | 33 |
| PS2123 | ΔprfA in frame | 33 |
| AG1468 | trpC2 pheA1 ΔspoOJ::spc | 13 |
| RB35 | trpC2 pheA1 Δsmc::kan | 6 |
| LP85 | ΔprfA::spc ΔthrC::(Pxyl-prfA xylR erm) | pLP79→PS2061 |
| LP88 | ΔprfA::spc ΔthrC::(Pxyl-prfA xylR erm) | pRDC19→PS2061 |
| LP89 | ΔthrC::(Pxyl-prfA xylR erm) | pLP79→PS832 |
| LP99 | Δsmc::kan | RB35→PS832 |
| LP101 | ΔprfA::spc ΔthrC::(Pxyl-prfA xylR erm) Δsmc::kan | RB35→LP85 |
| LP102 | ΔspoOJ::spc | AG1468→PS832 |
| LP105 | ΔprfA ΔspoOJ::spc | pNG7→LP102 |
| E. coli | ||
| PS1404 | BL21(DE3)/pLysS | 41 |
| PS2599 | BL21(DE3)/pLysS pET11a-dacC | 31 |
| PS2602 | BL21(DE3)/pLysS pET11a | 41 |
| PS2692 | BL21(DE3)/pLysS pET11a-dacC-NC | 31 |
| LP77 | BL21(DE3)/pLysS pET9d-prfA | pLP76→PS1404 |
a
B. subtilis was transformed as described previously (3), and transformants were selected on 2× SG agar plates at 37°C (all except RB35 transformants) or LB agar plates at 30°C (RB35 transformants) with appropriate antibiotics at the following concentrations: 100 μg of spectinomycin per ml, 10 μg of kanamycin per ml, or 0-5 μg of erythromycin per ml plus 12.5 μg of lincomycin per ml.