TABLE 1.
Bacterial strains and plasmids
| Designation | Relevant characteristic(s) | Source or reference |
|---|---|---|
| E. coli strains | ||
| JM109 | F′ ΔtraD36 lac1q Δ(lacZ)M15 proAB/recA1 endA1 gyrA96 thi hsdR17(rK− mK+) supE44 relA1 Δ(lac-proAB) | Lab stock |
| DH10B | mcrA Δ(mrr-hsdRMS-mcrBC) φ80dlacZΔM15 ΔlacX74 deoR recA1 endA1 araD139 Δ(ara, leu)7697 galU galK rpsL nupG | Lab stock |
| M. smegmatis strains | ||
| LR222 | Efficient plasmid transformation strain of mc26 | 2 |
| LUN1 | LR222 with Tn611 insertion in exochelin gene fxbC | 52 |
| LUN2 | LR222 with Tn611 insertion in exochelin gene exiT | 52 |
| LUN8 | LR222 with Tn611 insertion in fadD (mycobactin neg.) | This study |
| LUN9 | LR222 with Tn611 insertion in mbtE (mycobactin neg.) | This study |
| Plasmids | ||
| pCG79 | Kanr; temperature-sensitive, E. coli-mycobacterial shuttle plasmid containing Tn611 | 17 |
| pNBV1 | Hyr; E. coli-mycobacterial shuttle vector | 20 |
| pUC18 | Ampr; multiple cloning site vector | Lab stock |
| pCR2.1-TOPO | AT direct cloning vector for PCR products | Invitrogen |
| pCR4Blunt-TOPO | Blunt-end direct PCR cloning vector | Invitrogen |
| Litmus 28 | Ampr; multiple cloning site vector | NEBa |
| pARC6 | pUC18 with 4.4-kb SmaI fragment from LUN9 | This study |
| pWEN3 | pUC18 with 3.5-kb SacI fragment from LUN8 | This study |
| pML222 | pCG79 excised from LUN8 genome by EcoRI digestion | This study |
| pCR-Rv1344 | 3.3-kb PCR product from H37Rv cloned into pCR2.1-TOPO, encodes Rv1344-fadD33-fadE14 | This study |
| pML1344 | Litmus 28 with 3.3-kb EcoRI fragment from pCR-Rv1344 | This study |
| pMLNV1344 | pNBV1 with 3.3-kb HindIII-SpeI insert from pML1344 | This study |
| pCR4-Sm1344 | 3.6-kb PCR product from LR222 cloned into pCR4Blunt-TOPO, encodes acp-fadD-fadE | This study |
| pBLT1344 | pNBV1 with SnaBI insert from pCR4-Sm1344 cloned into the EcoRV site | This study |
a
New England Biolabs, Inc.