Table 1.
Strains, plasmids, and bacteriophages used in this study
| Strain, plasmid, or bacteriophage | Genotype or description | Application | Reference or source |
|---|---|---|---|
| Strains | |||
| E. coli DH5α | F− φ80lacZΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17 (rK− mK+) phoA supE44 λ−thi-1 gyrA96 relA1 | Lab collection | |
| E. coli TOP10 | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara leu)7697 galU galK rpsL (StrR) endA1 nupG | Invitrogen | |
| E. coli HB101 | supE44 hsdS20(rB− mB−) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 mtl-1 leuB6 thi-1 | Lab collection | |
| E. coli BL21(DE3)/pLysS | F−ompT hsdSB(rB− mB−) gal dcm (DE3)/pLysE; Camr | Novagen | |
| M. smegmatis mc2155 | Laboratory strain | ATCC 700084 | |
| M. tuberculosis H37Rv | Laboratory strain | ATCC 27294 | |
| TB193 | H37Rv ΔRv0081 | This study | |
| TB222 | H37Rv ΔRv0081 carrying pTZ1165 | This study | |
| TB228 | H37Rv ΔRv0081 carrying pTZ1111 and pTZ1161 | This study | |
| TB229 | H37Rv ΔRv0081 carrying pTZ1111 and pTZ1165 | This study | |
| Plasmids | |||
| pCR2.1-TOPO | 3.9-kb plasmid for cloning PCR products; Ampr Kanr | PCR cloning vector | Invitrogen |
| pET15b | Inducible His-tagged protein expression plasmid; Ampr | Overexpression | Lab collection |
| pJEM15 | E. coli-Mycobacterium shuttle vector for transcriptional fusion with lacZ; ampr | Reporter assay | 62 |
| pMV361 | Integrative protein expression vector in mycobacterial cell under hsp60 promoter; Kanr | Protein expression | 58 |
| pTZ229 | pET15b containing mprA coding sequence; Ampr | Overexpression | 70 |
| pTZ763 | pYUB854 containing dosR upstream and downstream regions; Hygr | dosR deletion | This study |
| pTZ765 | pET15b containing dosR coding sequence; Ampr | Overexpression | This study |
| pTZ1064 | pYUB854 containing mprA upstream and mprB downstream regions; Hygr | mprAB deletion | This study |
| pTZ1070 | pET15b containing Rv0081 coding sequence; Ampr | Overexpression | This study |
| pTZ1110 | pYUB854 containing Rv0081 upstream and downstream regions; Hygr | Rv0081 deletion | This study |
| pTZ1111 | pJEM15 containing Rv0081 upstream sequence; Kanr | Reporter assay | This study |
| pTZ1161 | pMV361 derivative with Kanr replaced by Hygr; Hygr | Protein expression | This study |
| pTZ1165 | pTZ1161 containing Rv0081 coding sequence; Hygr | Protein expression | This study |
| pYUB854 | 3.9-kb cosmid for generating gene deletions in Mycobacterium; carries Hygr cassette flanked by multiple cloning sites and λpac site; Hygr | Mycobacterium gene deletion vector | 4 |
| pYUB870 | 8.9-kb plasmid for unmarking mutations in Mycobacterium; carries tnpR and sacB cassettes; Kanr | Mycobacterium gene deletion unmarking vector | 4 |
| Mycobacteriophages | |||
| phAE87 | Derivative of conditionally replicating mycobacteriophage PH101(Ts) | 4 | |
| phTZ763 | phAE87 derivative containing pTZ763 | dosR deletion | This study |
| phTZ1064 | phAE87 derivative containing pTZ1064 | mprAB deletion | This study |
| phTZ1110 | phAE87 derivative containing pTZ1110 | Rv0081 deletion | This study |