TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant genotype or featuresa | Source or referenceb |
|---|---|---|
| Strains | ||
| M. tuberculosis H37Rv | Virulent laboratory strain | ATCC |
| E. coli | ||
| JM109 | e14− (McrA−) recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 Δ(lac-proAB) | Stratagene |
| T7 Express lysY | miniF lysY (Camr)/fhuA2 lacZ::T7 gene1 [lon] ompT gal sulA11 R(mcr-73::mini-Tn10-Tets)2 [dcm] R(zgb-210::Tn10-Tets) endA1 Δ(mcrC-mrr)114::IS10 | NEB |
| SK140 | E. coli T7 Express lysY with pET28a(+) | This study |
| SK141 | E. coli T7 Express lysY with pET28::relB | This study |
| M. smegmatis mc2155 | ept-1 | 45 |
| LIX22 | mc2155 with pJEM15 | This study |
| LIX23 | mc2155 with pJEM15::PrelBE | This study |
| LIX24 | mc2155 with pJEM15::PrelBE-relB | This study |
| LIX25 | mc2155 with pJEM15::PrelBE-relBE | This study |
| LIX26 | mc2155 with pJEM15::PrelFG | This study |
| LIX27 | mc2155 with pJEM15::PrelFG-relF | This study |
| LIX28 | mc2155 with pJEM15::PrelFG-relFG | This study |
| LIX29 | mc2155 with pJEM15::PrelJK | This study |
| LIX30 | mc2155 with pJEM15::PrelJK-relJ | This study |
| LIX31 | mc2155 with pJEM15::PrelJK-relJK | This study |
| LIX32 | mc2155 with pYA1611 | This study |
| LIX33 | mc2155 with pYA1611::relE | This study |
| LIX34 | mc2155 with pYA1611::relBE | This study |
| LIX35 | mc2155 with pYA1611::relG | This study |
| LIX36 | mc2155 with pYA1611::relFG | This study |
| LIX37 | mc2155 with pYA1611::relK | This study |
| LIX38 | mc2155 with pYA1611::relJK | This study |
| Plasmids | ||
| pJEM15 | Kanr, pAL5000 replicon, cII-lacZ | 47 |
| pJEM15::PrelBE | PrelBE in ApaI-SphI sites of pJEM15 | This study |
| pJEM15::PrelBErelB | PrelBE + relB in ApaI-SphI sites of pJEM15 | This study |
| pJEM15::PrelBErelBE | PrelBE + relBE in ApaI-SphI sites of pJEM15 | This study |
| pJEM15::PrelFG | PrelFG in ApaI-SphI sites of pJEM15 | This study |
| pJEM15::PrelFGrelF | PrelFG + relF in ApaI-SphI sites of pJEM15 | This study |
| pJEM15::PrelFGrelFG | PrelFG + relFG in ApaI-SphI sites of pJEM15 | This study |
| pJEM15::PrelJK | PrelJK in ApaI-BamHI sites of pJEM15 | This study |
| pJEM15::PrelJKrelJ | PrelJK + relJ in ApaI-BamHI sites of pJEM15 | This study |
| pJEM15::PrelJKrelJK | PrelJK + relJK in ApaI-BamI sites of pJEM15 | This study |
| pSE100 | Hygr, Pmyc1tetO promoter, Myc ori | 19 |
| pYA1611 | tetR gene in NotI site of pSE100 | This study |
| pYA1611::relE | relE gene in PstI site of pYA1611 | This study |
| pYA1611::relBE | relBE gene in PstI site of pYA1611 | This study |
| pYA1611::relG | relG gene in PstI site of pYA1611 | This study |
| pYA1611::relFG | relFG gene in PstI site of pYA1611 | This study |
| pYA1611::relK | relK gene in PstI site of pYA1611 | This study |
| pYA1611::relJK | relJK gene in PstI site of pYA1611 | This study |
| pSH276 | rv0195 gene in pQE40ΔDHFR | 20 |
| pSH337 | clpC gene in pGEM-T | 20 |
| pSH317 | devR gene in pQE40ΔDHFR | 20 |
| pSH281 | rv2027c gene in pQE40ΔDHFR | 20 |
| pSH334 | regX3 gene in pZERO/Km | 20 |
| pGEM-T | Ampr, 3′-T overhang for PCR product ligation | Promega |
| pGEM::relE | relE gene in pGEM-T | This study |
| pGEM::relG | relG gene in pGEM-T | This study |
| pGEM::relK | relK gene in pGEM-T | This study |
| pGEM::relB | relB gene in pGEM-T | This study |
| pGEM::relF | relF gene in pGEM-T | This study |
| pGEM::relJ | relJ gene in pGEM-T | This study |
| pGEX-4T-2 | GST fusion expression vector | GE Healthcare |
| pGEX::relB | relB in EcoRI-XhoI sites of pGEX-4T-2 | This study |
| pGEX::relE | relE in EcoRI-XhoI sites of pGEX-4T-2 | This study |
| pET28a(+) | Kanr, T7 lac promoter-operator, N-terminal His6 | Novagen |
| pET28::relB | relB in NcoI-BamHI sites of pET28a(+) | This study |
| pUAB100 | Hygr, oriM oriE hsp60-GCN4-Gly10-mDHFR [F1,2] | 44 |
| pUAB200 | Kanr, attP int hsp60-GCN4-Gly10-mDHFR [F3] | 44 |
| pUAB300 | Hygr, oriM oriE hsp60-mDHFR [F1,2]-Gly10-MCS | 44 |
| pUAB400 | Kanr, attP int hsp60-mDHFR [F3]-Gly10-MCS | 44 |
| pUAB100::relB | relB in BamHI-ClaI sites of pUAB100, creating RelB[F1,2]-C | This study |
| pUAB200::relE | relE in MfeI-ClaI sites of pUAB200, creating RelE[F3]-C | This study |
| pUAB300::relB | relB in BamHI-ClaI sites of pUAB300, creating RelB[F1,2]-N | This study |
| pUAB400::relE | relE in MfeI-ClaI sites of pUAB400, creating RelE[F3]-N | This study |
a
GCN4 is the S. cerevisiae leucine-zipper sequence. MCS, multiple cloning site.
b
ATCC, American Type Culture Collection; NEB, New England Biolabs.