TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Genotype or description | Source or reference |
|---|---|---|
| Strains | ||
| Escherichia coli | ||
| DH5α | K-12 DH5α F− φ80dΔlacZM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17 glnV44 thi-1 gyrA96 relA1 | Gibco-BRL |
| DH5α λpir | K-12 DH5α F− φ80dΔlacZM15 Δ (lacZYA-argF)U169 deoR recA1 endA1 hsdR17 glnV44 thi-1 gyrA96 relA λpir | W. R. Jacobs, Jr. |
| Mycobacterium | ||
| mc2155 | M. smegmatis ept-1 | 48 |
| H37Rv | Virulent M. tuberculosis | 3 |
| PM638 | H37Rv ΔblaC | 17 |
| MBTB126 | PM638 carrying integrating vector pJES137 | This study |
| MBTB204 | PM638 with a Tn′blaTEM-1 insertion in mce1A (Rv0169) at amino acid 449 of 455 | This study |
| MBTB190 | PM638 with a Tn′blaTEM-1 insertion in mce1B (Rv0170) at amino acid 223 of 337 | This study |
| MBTB183 | PM638 with a Tn′blaTEM-1 insertion in rv0199 at amino acid 74 of 220 | This study |
| MBTB156 | PM638 with a Tn′blaTEM-1 insertion in mce2F (Rv0594) at amino acid 476 of 517 | This study |
| MBTB381 | PM638 with a Tn′blaTEM-1 insertion in ctaC (Rv2200c) at amino acid 351 of 364 | This study |
| MBTB301 | PM638 with a Tn′blaTEM-1 insertion in lppX (Rv2945c) at amino acid 82 of 234 | This study |
| Plasmids | ||
| pCR2.1 | bla aph ColE1; TA cloning vector | Invitrogen |
| pMV261.kan | aph Phsp60oriM ColE1; multicopy mycobacterial shuttle plasmid | 51 |
| pJES102 | aph Phsp60-′blaTEM-1 (E. coli) oriM ColE1; ′blaTEM-1 from pCR2.1 cloned into pMV261.kan | 34 |
| pSH1 | TnHimar1 oriR6Kg Hygr; shuttle vector carrying the Himar1 transposon | 27 |
| pJES123 | hygR oriR6K ′blaTEM-1; intermediate cloning vector | This study |
| pJES124 | hygR oriR6K ColE1 aph Tn′blaTEM-1 Tnpmar λcos; shuttle vector carrying Tn′blaTEM-1 | This study |
| pJES137 | hyg int attP ColE1 Phsp60 ′blaTEM-1; integrating hygromycin-marked vector with constitutively expressed ′BlaTEM-1 | This study |
| pJES153 | aph Phsp60-mce2F oriE oriM; multicopy kanamycin-marked vector with constitutively expressed mce2F | This study |
| pJES178 | aph Phsp60-rv0199 oriE oriM; multicopy kanamycin-marked vector with constitutively expressed Rv0199 | This study |