Table 2.
Plasmids used in this study
| Plasmid | Characteristics | Source or reference |
|---|---|---|
| pJL37 | Shuttle plasmid, KanR, oriM, oriE, M. bovis BCG hsp60 promoter | Lewis and Hatfull (2000) |
| pMP1 | pJL37 containing MSMEG3721 CDS with the putative SP (NdeI-HindIII) | This work |
| pMP2 | pJL37 containing MSMEG3721 CDS without the putative SP (NdeI-HindIII) | This work |
| pLAM12 | Derivative of pJL37, BCG hsp60 promoter was replaced with an acetamidase promoter from pSD26 | Lab collection |
| pMP3 | pLAM12 containing MSMEG3721 CDS with the putative SP (NdeI-HindIII) | This work |
| pMH94 | Mycobacteria integrative vector, KanR, L5 attP and integrase | Lee et al. (1991) |
| pMP4 | pMH94 containing MSMEG3721 CDS with the putative SP under control of acetamidase promoter from pMP3 (XbaI-NheI) | This work |
| pYUB854 | Cosmid, oriE, HygR, res-hyg-res | Bardarov et al. (2002) |
| pMP5 | pYUB854 containing 5′ and 3′ region of MSMEG3721 and 600 bp of upstream and downstream DNA (XbaI-XmaI and XhoI-BstBI) | This work |
| pMP6 | pYUB854 containing 5′ and 3′ region of MSMEG0642 and 600 bp of upstream and downstream DNA (KpnI-XbaI and XhoI-BglII) | This work |
| pMP7 | pYUB854 containing 5′ and 3′ region of MSMEG6076 and 600 bp of upstream and downstream DNA (AflII-XbaI and HindIII-XhoI) | This work |
| pPG1 | Derivative of pJL37 lacking the BCG hsp60 promoter | Ghosh et al. (2006) |
| pMP8 | pPG1 containing MSMEG3721 gene with the putative promoter and transcription terminator sequences (PCR fragment cloned in BsaAI) | This work |
| pMP9 | pPG1 containing MSMEG0642 gene with the putative promoter and transcription terminator sequences (PCR fragment cloned in BsaAI) | This work |
| pET-28a | Cloning/inducible expression vector, KanR, N-terminal His Tag | Novagen |
| pMP10 | pET-28a containing MSMEG3721 CDS without the putative SP (NdeI-HindIII) | This work |
| pMP11 | pET-28a containing MSMEG0642 CDS without the putative SP (NdeI-HindIII) | This work |
| pMP12 | pET-28a containing MSMEG3721 TM4 Tmp mt3 (hybrid protein) (NdeI-HindIII) | This work |
| phAE87 | pH101::pYUB328 that fails to replicate at 37°C. PH101 is a ts mutant of TM4, AmpR | Bardarov et al. (1997) |
| phAE87Δ17mt3 | pHAE87Δ mt3 in gene 17 | This work |
| phAE8717mt3W4G | phAE87 with a single base change T-G in gene 17 that generates a W4G substitution in Tmp mt3 | This work |
| pYUB556 | Cosmid, KanR, oriE and FFlux under control of BCG hsp60 promoter | Carriere et al. (1997) |
| pJV24 | KanR, oriM, Che9c gp59–62 under control of acetamidase promoter | J.C. van Kessel and G.F. Hatfull (submitted) |
| pGH542 | TetR, oriM, γδ resolvase under control of BCG hsp60 promoter | Lab collection |