TABLE 1.
Plasmids used in this study
| Plasmid | Relevant features | Source or reference |
|---|---|---|
| pFPV27 | E. coli-Mycobacterium shuttle plasmid with promoterless GFP gene, Kanr | 37 |
| p1738 | GFP reporter plasmid containing Rv1738 promoter, Kanr | 16 |
| pJFR19 | E. coli-Mycobacterium integrating shuttle plasmid with 3-kb acetamidase promoter, Hygr | 22 |
| pUAB100 | E. coli-Mycobacterium shuttle plasmid expressing GCN4-mDHFR F[1,2] under hsp60 promoter, Hygr | 23 |
| pUAB200 | E. coli-Mycobacterium shuttle plasmid expressing GCN4-mDHFR F[3] under hsp60 promoter, Kanr | 23 |
| pUS Phsp60 | pJFR19 containing constitutive hsp60 promoter cloned in NdeI and BstBI sites | This study |
| pUS Phsp60 devRwt | pUS Phsp60 containing devR wild type (cloned in NdeI sites); DevR is expressed from the constitutive hsp60 promoter | This study |
| pUS Phsp60 devRmut | pUS Phsp60 containing devR mutant variant generated by site-directed mutagenesis; individual mutant DevR proteins (D152A, E154A, etc.) are expressed from the constitutive hsp60 promoter | This study |
| pUS-mut DevR | pGEX4T1 overexpressing DevR E154A or other mutant DevR proteinsa | This study |
| pAV-FLdevR100 | pUAB100 plasmid containing devR (cloned in BamHI and ClaI sites); DevR-mDHFR F[1,2] protein is expressed from the constitutive hsp60 promoter, Hygr | This study |
| pAV-E154A | pAV-FLdevR100 derivative expressing DevR with E154A mutation | This study |
| pAV-E178A | pAV-FLdevR100 derivative expressing DevR with E178A mutation | This study |
| pVS-SigA200 | pUAB200 plasmid containing SigA (cloned in MfeI and ClaI sites); SigA-mDHFR F[3] is expressed from the constitutive hsp60 promoter, Kanr | This study |
a
WT and all DevR mutant proteins are expressed with an N-terminal GST tag.