Table 1.
Bacterial strains and plasmids used in this study
| Bacterial strain/plasmid | Relevant genotype or characteristics | Reference or source |
|---|---|---|
| M. bovis BCG | Pasteur strain | VLA Weybridge, UK |
| M. bovis AF2122/97 | Virulent isolate from a cow in Cornwall, UK | VLA Weybridge, UK |
| M. tuberculosis H37Rv | Virulent strain isolated in 1905 | VLA Weybridge, UK |
| M. smegmatis mc2155 | Mutant with high plasmid transformation efficiency | Professor W. R. Jacobs, AECOM, NY, USA |
| M. bovis BCG Δbcg3145 | Knockout strain of M. bovis BCG Pasteur with bcg3145 gene replaced by hygromycin cassette | This study |
| E. coli DH5α | General cloning strain | Invitrogen |
| E. coli BL21(DE3) | Strain used for protein expression | Invitrogen |
| E. coli HB101 | General cloning strain | Invitrogen |
| E. coli XL1-Blue | General cloning strain | Invitrogen |
| pUC19 | General cloning vector | VLA Weybridge |
| pUC3124 | pUC19 containing rv3124 | This study |
| pSM96 | Mycobacterial expression vector with hsp60 promoter | VLA Weybridge, UK (Wooff et al., 2002) |
| pB1 | rv3124 in pSM96 | This study |
| pB3 | bcg3145 in pSM96 | This study |
| MBPpET | MBP–His fusion vector used in protein purification | Vircell |
| pYUB854 | Cosmid vector used for mutant construction; contains the hygromycin cassette and a multiple cloning site | Bardarov et al. (2002) |
| p3145KO | Cosmid used in the bcg3145 knockout containing the gene flanking regions into pYUB854 | This study |
| pSM128 | Mycobacteriophage L5-based vector carrying a lacZ reporter | Dussurget et al. (1999) |
| pB31 | 190 bp upstream of rv3109 in pSM128 | This study |
| pB32 | 116 bp upstream of rv3109 in pSM128 | This study |
| pB33 | 45 bp upstream of rv3109 in pSM128 | This study |
| pB10 | G90A mutation in the DNA-binding domain | This study |
| pB11 | Y91A mutation in the DNA-binding domain | This study |
| pB15 | E159I mutation in the BTA domain | This study |
| pB17 | E159W mutation in the BTA domain | This study |
| pB23 | E159D mutation in the BTA domain | This study |
| pB25 | E159Q mutation in the BTA domain | This study |