Table 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristics | Source or reference |
|---|---|---|
| Strains | ||
| M. tuberculosisstrain | ||
| 1424 | M. tuberculosisH37Rv derivative; Strr | 6 |
| E. colistrains | ||
| DH5α | Invitrogen | |
| BL21(DE3) | Invitrogen | |
| Plasmids | ||
| pBS-Int | pBluescript carrying the L5 integrase gene | 32 |
| pKP186 | Cloning vector carrying L5 attPsite; Kanr | 24 |
| pKP203 | Cloning vector carrying L5 attPsite; Gmr | 24 |
| pAW24 | uvrD2in pKP186 | This study |
| pAW26 | uvrD2in pKP203 | This study |
| pAW27 | uvrD21-600in pKP203 | This study |
| pAW28 | uvrD21-640in pKP203 | This study |
| pAW29 | uvrD2(Q266R) in pKP203 | This study |
| pAW30 | uvrD2(E508A) in pKP203 | This study |
| pMCS5-rpsL-hyg | rpsL-containing allele-exchange vector | 35 |
| pUVRD2-targ | pMCS5-rpsL-hyg containing PCR-amplified 5′ (1,316 bp) and 3′ (1,225 bp) regions surrounding uvrD2 | This study |
| pET-28 | Expression vector, contains histidine tag | Novagen |
| pET28-UvrD2 | uvrD2with C-terminal histidine tag and T7 promoter | This study |
| pET28-Q266R | uvrD2(Q266R) with C-terminal histidine tag | This study |
| pET28-E508A | uvrD2(E508A) with C-terminal histidine tag | This study |