TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Characteristicsa | Reference or source |
|---|---|---|
| Strains | ||
| M. vaccae JOB5 | Wild type, MTBE-degrading strain; G+ | DSMZ |
| M. smegmatis mc2-155 | Wild type, used for gene expression; G+ | ATCC |
| mc2-155-pJV53 | mc2-155 transformed with pJV53; Kmr | This study |
| mc2-155-pJV53-prmABCD | mc2-155 transformed with pJV53-prmABCD; Kmr | This study |
| E. coli | ||
| DH5α | supE44 lacU169(80dlacZΔM15) hsdR17 recA1 endA1 gyrA96 Δthi relA1 | Tsingke |
| BL21(DE3) | Chaperonin gene groELS integrated in genome | 37 |
| Rosetta(DE3) | F- ompT hsdSB (rB−, mB−) gal dcm(DE3) pRARE; CamR | TransGen |
| Rosetta(DE3)-pET28a | Rosetta(DE3) transformed with pET28a; Kmr | This study |
| Rosetta (DE3)-pET28a-prmABCD | Rosetta(DE3) transformed with pET28a-prmABCD; Kmr | This study |
| Rosetta(DE3)-pET28a-prm-A-3*FLAG | Rosetta(DE3) transformed with pET28a-prm-A-3*FLAG; Kmr | This study |
| Rosetta(DE3)-pET28a-prm-B-3*FLAG | Rosetta(DE3) transformed with pET28a-prm-B-3*FLAG; Kmr | This study |
| Rosetta(DE3)-pET28a-prm-C-3*FLAG | Rosetta(DE3) transformed with pET28a-prm-C-3*FLAG; Kmr | This study |
| Rosetta(DE3)-pET28a-prm-D-3*FLAG | Rosetta(DE3) transformed with pET28a-prm-D-3*FLAG; Kmr | This study |
| Rosetta(DE3)-pET28a-prmA | Rosetta(DE3) transformed with pET28a-prmA; Kmr | This study |
| Rosetta(DE3)-pET28a-prmB | Rosetta(DE3) transformed with pET28a-prmB; Kmr | This study |
| Rosetta(DE3)-pET28a-prmC | Rosetta(DE3) transformed with pET28a-prmC; Kmr | This study |
| Rosetta(DE3)-pET28a-prmD | Rosetta(DE3) transformed with pET28a-prmD; Kmr | This study |
| Rosetta(DE3)-pET28a-prmAB-His | Rosetta(DE3) transformed with pET28a-prmAB-His; Kmr | This study |
| Rosetta(DE3)-pET28a-prmC-His | Rosetta(DE3) transformed with pET28a-prmC-His; Kmr | This study |
| Rosetta(DE3)-pET28a-prmD-His | Rosetta(DE3) transformed with pET28a-prmD-His; Kmr | This study |
| Rosetta(DE3)-pET28a-prmABC | Rosetta(DE3) transformed with pET28a-prmABC-His; Kmr | This study |
| Rosetta(DE3)-pET28a-prmABD | Rosetta(DE3) transformed with pET28a-prmABD-His; Kmr | This study |
| Rosetta(DE3)-pET28a-prmBCD | Rosetta(DE3) transformed with pET28a-prmBCD-His; Kmr | This study |
| Rosetta(DE3)-pET28a-prmAB | Rosetta(DE3) transformed with pET28a-prmAB-His; Kmr | This study |
| Rosetta(DE3)-pET28a-prm-I84V | Rosetta(DE3) transformed with pET28a-prm-I84V; Kmr | This study |
| Rosetta(DE3)-pET28a-prm-G95A | Rosetta(DE3) transformed with pET28a-prm-G95A; Kmr | This study |
| Rosetta(DE3)-pET28a-prm-P165A | Rosetta(DE3) transformed with pET28a-prm-P165A; Kmr | This study |
| Rosetta(DE3)-pET28a-prm-A269S | Rosetta(DE3) transformed with pET28a-prm-A269S; Kmr | This study |
| Rosetta(DE3)-pET28a-prm-V270T | Rosetta(DE3) transformed with pET28a-prm-V270T; Kmr | This study |
| Rosetta(DE3)-pET28a-prm-Q288E | Rosetta(DE3) transformed with pET28a-prm-Q288E; Kmr | This study |
| BL21(DE3)-pET28a | BL21DE3) transformed with pET28a; Kmr | This study |
| BL21(DE3)-pET28a-prmABCD | BL21(DE3) transformed with pET28a-prm; Kmr | This study |
| Plasmids | ||
| pET-28a | Gene expression vector; Kmr | Novagen |
| pET28a-prmABCD | Vector for inducible expression of prmABCD; Kmr | This study |
| pET28a-prm-A-3*FLAG | Vector for inducible expression of prm-A-3*FLAG; Kmr | This study |
| pET28a-prm-B-3*FLAG | Vector for inducible expression of prm-B-3*FLAG; Kmr | This study |
| pET28a-prm-C-3*FLAG | Vector for inducible expression of prm-C-3*FLAG; Kmr | This study |
| pET28a-prm-D-3*FLAG | Vector for inducible expression of prm-D-3*FLAG; Kmr | This study |
| pET28a-prmA | Vector for inducible expression of prmA; Kmr | This study |
| pET28a-prmB | Vector for inducible expression of prmB; Kmr | This study |
| pET28a-prmC | Vector for inducible expression of prmC; Kmr | This study |
| pET28a-prmD | Vector for inducible expression of prmD; Kmr | This study |
| pET28a-prmAB-His | Vector for inducible expression of prmAB-His; Kmr | This study |
| pET28a-prmC-His | Vector for inducible expression of prmC-His; Kmr | This study |
| pET28a-prmD-His | Vector for inducible expression of prmD-His; Kmr | This study |
| pET28a-prmABC | Vector for inducible expression of prmABC; Kmr | This study |
| pET28a-prmABD | Vector for inducible expression of prmABD; Kmr | This study |
| pET28a-prmBCD | Vector for inducible expression of prmBCD; Kmr | This study |
| pET28a-prmAB | Vector for inducible expression of prmAB; Kmr | This study |
| pET28a-prm-I84V | Vector for inducible expression of prm-I84V; Kmr | This study |
| pET28a-prm-G95A | Vector for inducible expression of prm-G95A; Kmr | This study |
| pET28a-prm-P165A | Vector for inducible expression of prm-P165A; Kmr | This study |
| pET28a-prm-A269S | Vector for inducible expression of prm-A269S; Kmr | This study |
| pET28a-prm-V270T | Vector for inducible expression of prm-V270T; Kmr | This study |
| pET28a-prm-Q288E | Vector for inducible expression of prm-Q288E; Kmr | This study |
| pJV53 | Gene expression vector; Kmr | This study |
| pJV53-prmABCD | Vector for inducible expression of prmABCD; Kmr | This study |
a
Kmr, kanamycin resistance.