TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristica | Reference |
|---|---|---|
| Strains | ||
| E. coli DH5α | F−recA1 hsdR17 thi-1 gyrA96 supE44 endA1 relA1 recA1 deoR Δ(lacZYA-argF)U169 (φ80 lacZ ΔM15) | 6 |
| M. bovis BCG | Pasteur strain | ATCC 35734 |
| M. smegmatis | mc26 1-2C | 47 |
| Plasmids | ||
| pUC19 | High-copy-number E. coli plasmid, Apr | 46 |
| pNBV1 | E. coli-mycobacterial shuttle plasmid, Hyr | 22 |
| pYZ99 | pUC18 containing sigF on a 2.8-kb BamHI fragment from M. tuberculosis H37Rv, Apr | 12 |
| pLZK82 | pUC19 containing lacZ-Kmr on a 4.4-kb BamHI fragment | 5 |
| pMH5 | Integration-proficient pUC119 derivative containing a 4.9-kb insert of the phage L5 genome, Kmr | 27 |
| pIJV1 | pUC19- and pMH5-derived integrating E. coli-mycobacterial shuttle plasmid | 15 |
| pCK2819-P | pUC19 containing sigF on a 2.8-kb PstI fragment | This study |
| pCK2819-EB | pUC19 containing sigF with an additional BamHI site adjacent to EcoRI on a 2.8-kb PstI fragment | This study |
| pCK0221 | pUC19 containing frameshifted sigF::lacZ on a 7.6-kb SapI-XbaI fragment, Kmr Apr | This study |
| pCK2295 | pUC19 containing in-frame sigF::lacZ on a 7.6-kb SapI-XbaI SapI-XbaI fragment, Kmr Apr | This study |
| pCK3127 | pNBV1 containing in-frame sigF::lacZ on a 7.6-kb SapI-XbaI fragment, Kmr Hyr | This study |
| pCK1107 | pNBV1 containing frameshifted sigF::lacZ on a 7.6-kb SapI-XbaI fragment, Kmr Hyr | This study |
| pCK3215 | pIJV1 containing in-frame sigF::lacZ on a 7.6-kb SapI-XbaI fragment, Kmr, integrating | This study |
| pGS16 | Integrating E. coli-mycobacterial shuttle plasmid containing FFlux Kmr Apr | 24 |
a
Abbreviations: Ap, ampicillin; Hy, hygromycin; Km, kanamycin.