TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Description | Source or reference |
|---|---|---|
| Strains | ||
| M. smegmatis | ||
| mc2155 | High-frequency transformation mutant of M. smegmatis ATCC 607 | 32 |
| pzaA::aph | pzaA knockout mutant of mc2155; Kmr | 2 |
| M. tuberculosis | ||
| H37Rv | Laboratory strain (ATCC 27294) | Laboratory collection |
| pncA::hyg | pncA knockout mutant of H37Rv; Hygr | This work |
| Plasmids | ||
| pGEM3Z(+)f | E. coli cloning vector; Apr | Promega |
| pET15b | E. coli expression vector; Apr | Novagen |
| pMAL-c2 | E. coli expression vector; Apr | New England Biolabs |
| pHINT | E. coli-Mycobacterium integrating shuttle vector; Hygr Apr | 21 |
| pAINT | pHINT derivative with hyg replaced by aph; Kmr Apr | This work |
| pOLYG | E. coli-mycobacterium replicating shuttle vector; Hygr oriM | 21 |
| Pac4 | Cosmid from pYUB328::H37Rv library carrying the pncA gene | This work |
| pGncA | pGEM3Z(+)f derivative carrying pncA on a 4.67-kb Asp718 fragment | This work |
| pGncAh | pGncA derivative carrying pncA insertionally inactivated by hyg (pncA::hyg) | This work |
| pGncAKO | pGncAh derivative carrying hsp60-sacB | This work |
| pHncA | pHINT derivative carrying pncA on a PstI fragment | This work |
| pOncA | pOLYG derivative carrying pncA on a PstI fragment | This work |
| pOppncA | pOLYG derivative carrying pncA with consensus ribosome binding site under control of the pzaA promoter | This work |
| pTncA | pET15b derivative carrying pncA | 2 |
| pAIncA | pAINT derivative carrying pncA on a 4.67-kb Asp718 fragment | This work |
| pMncA | pMAL-c2 derivative carrying malE-pncA translational fusion | This work |
| pGam | pGEM3Z(+)f derivative carrying pzaA with upstream sequence | 2 |
| pOam | pOLYG derivative carrying pzaA with upstream sequence | 2 |
| pAIam | pAINT derivative carrying pzaA with upstream sequence | This work |
| pTam | pET15b derivative carrying pzaA | 2 |