TABLE 1.
Strains, plasmids, and bacteriophages used in this study
| Strain, plasmid, or bacteriophage | Genotype or description | Application | Reference or source |
|---|---|---|---|
| Strains | |||
| M. tuberculosis H37Rv | Laboratory strain | ATCC 27294 | |
| M. smegmatis mc2155 | Laboratory strain | ATCC 700084 | |
| E. coli DH5α | [λ− φ80dlacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK− mK−) supE44 thi-1 gyrA relA1] | Cloning | Laboratory collection |
| E. coli TOP10 | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara leu)7697 galU galK rpsL (StrR) endA1 nupG | Cloning | Invitrogen |
| E. coli HB101 | supE44 hsdS20(rB− mB−) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 mtl-1 leuB6 thi-1 | E. coli transduction host | Laboratory collection |
| E. coli BL21(DE3)/pLysS | F−ompT hsdSB(rB− mB−) gal dcm (DE3)/pLysE (Camr) | Protein overexpression | Novagen |
| Plasmids | |||
| pCR2.1-TOPO | 3.9-kb plasmid for cloning PCR products; Ampr Kanr | PCR cloning vector | Invitrogen |
| pET-24b | 5.3-kb plasmid allowing C-terminal protein fusion to His6; Kanr | Protein overexpression vector | Novagen |
| pMV306 | 3.9-kb plasmid for single-copy integration into mycobacterial chromosome; Kanr | Complementation vector | Laboratory collection |
| pTZ557 | pYUB854 containing pepD upstream and downstream regions; Hygr | pepD deletion | This study |
| pTZ612 | pYUB854 containing sigE upstream and downstream regions; Hygr | sigE deletion | This study |
| pTZ758 | pET-24b containing pepDΔTM coding sequence; Kanr | Overexpression | This study |
| pTZ775 | pET-24b containing pepDΔTMΔPDZ coding sequence; Kanr | Overexpression | This study |
| pTZ792 | pMV306 containing pepD and moaB2 coding sequences; Kanr | Complementation | This study |
| pTZ797 | pET-24b containing pepDΔTMS317A coding sequence; Kanr | Overexpression | This study |
| pTZ844 | pYUB854 containing mprA upstream and mprB downstream regions; Hygr | mprAB deletion | This study |
| pTZ898 | pYUB854 containing sacB coding sequence; Hygr | Mycobacterium gene deletion vector | This study |
| pTZ903 | pMV306 containing pepDS317A and moaB2 coding sequences; Kanr | Complementation | This study |
| pTZ947 | pTZ898 containing mprAB upstream and downstream regions; Hygr | mprAB deletion | This study |
| pTZ949 | pTZ898 containing pepD upstream and downstream regions; Hygr | pepD deletion | This study |
| pTZ1065 | pMV306 containing sigE coding sequence; Kanr | Complementation | This study |
| pYUB854 | 3.9-kb cosmid for generating gene deletions in Mycobacterium. Carries a Hygr cassette flanked by multiple cloning sites and a λpac site; Hygr | Mycobacterium gene deletion vector | 5 |
| pYUB870 | 8.9-kb plasmid for unmarking mutations in Mycobacterium. Carries the tnpR and sacB cassettes; Kanr | Mycobacterium gene deletion unmarking vector | 5 |
| Mycobacteriophages | |||
| phAE87 | Derivative of conditionally replicating mycobacteriophage PH101(Ts) | 5 | |
| phTZ557 | phAE87 derivative containing pTZ557 | pepD deletion | This study |
| phTZ612 | phAE87 derivative containing pTZ612 | sigE deletion | This study |
| phTZ844 | phAE87 derivative containing pTZ844 | mprAB deletion | This study |