TABLE 1.
Bacterial strains and plasmids used in this study
| Relevant characteristicsa | Refs. | |
|---|---|---|
| E. coli DH5αF′ | F'/hsdR17, recA1, gyrA | (39) |
| E. coli BL21(DE3) | F-, ompl, hsdSB (r-B m-B) | (40) |
| P. putida DOT-T1E | phototroph, Tol+ (tod pathway) | (41) |
| P. putida DOT-T1EtodST | DOT-T1E, todST::Km, Tol-, containing pMlR66, GmR | (14) |
| pET-28b(+) | KmR, protein expression vector | (Novagen) |
| pMIR77 | TcR, PtodX::'lacZ inserted in pMP220 | (14) |
| pMIR66 | GmR, containing the todST genes inserted in pBBR1MCS-5 | (14) |
| pTodS | pET28b (+) containing todS gene | (15) |
| pNTodS | pET28b (+) containing todS sequence 1-1752 (amino acids 1-584, NTodS-long) | (15) |
| pCTodS | pET28b (+) containing todS sequence 1356-2934 (amino acids 452-978, CTodS-long) | (15) |
| pHK1 | pET28b (+) containing todS sequence 49-1347 (amino acids 16-449, NTodS-short) | This work |
| pRRR | pET28b (+) containing todS sequence 1348-1755 (amino acids 450-585, MTodS) | This work |
| pHK2 | pET28b (+) containing todS sequence 1756-2928 (amino acid 586-976, CTodS-short) | This work |
| pAB1 | pTodS containing the mutation H190A | This work |
| pAB2 | pTodS containing the mutation D500A | This work |
| pAB3 | pTodS containing the mutation H760A | This work |
| pAB4 | pMIR66 containing the mutation H190A | This work |
| pAB5 | pMIR66 containing the mutation D500A | This work |
| pAB6 | pMIR66 containing the mutation H760A | This work |
| pJLTodT | pET28b containing the todT gene | (15) |
| pTodTD57A | pJLTodT containing the mutation D57A | (15) |
a
Abbreviations: GmR, KmR,TcR resistance to gentamicin, kanamycin, and tetracycline, respectively. Tol+/Tol- indicates that the strain grows or fails to grow on toluene, respectively.