TABLE 1.
Bacterial strains and plasmids used in this study
| Bacterial strain or plasmid | Relevant characteristics | Source or reference |
|---|---|---|
| Bacterial strains | ||
| Pseudomonas sp. | ||
| HI-70 | Wild type, grows on cyclododecanol or cyclododecanone | This study |
| HI-70MB | cpdB::lacZ, Kmr; no growth on cyclododecanol or cyclododecanone | This study |
| E. coli | ||
| BL21(DE3) | F−ompT hsdSB(rB− mB−) gal dcm | Novagen |
| DH5α | supE44 thi-1 recA1 hsdR17 endA1 gyrA (Nalr) Δ(lacIZYA-argF)U169 deoR [φ80dlacΔ(lacZ)M15] | 49 |
| S17-1 | recA pro thi hsdR RP4-2-Tc::Mu-Km::Tn7 Tra+ Tpr Smr | 54 |
| XL1-Blue | recA1 endA1 gyrA96 thi hsdR17 supE44 relA1 [F′ lacIqZM15 Tn10(Tetr)] | 49 |
| Plasmids | ||
| pARO190 | Mobilizable vector; Apr | 45 |
| pARO-cpdB | pARO190 with a 2.9-kb EcoRV fragment containing cpdB from pCD200 | This study |
| pARO-cpdB::lacZ | cpdB::lacZ-Kmr; SalI-excised lacZ-Kmr cassette from pKOK6.1 in XhoI site of pARO-cpdB | This study |
| pKOK6.1 | lacZ-Kmr cassette | 30 |
| pSD80 | Expression vector with tac promoter; Apr | 55 |
| pUC19 | Cloning vector with lac promoter; Apr | 71 |
| pCD200 | pUC19 with a 4.2-kb BclI fragment from Pseudomonas sp. strain HI-70; Apr | This study |
| pCD201 | 1.8-kb EcoRIa fragment containing cpdB in pSD80; Apr | This study |
| pCpdBdelN | Deletion of CpdB N-terminal 54 amino acids in pCD201 | This study |
| pCpdBdel7 | Deletion of internal 7 amino acids in CpdB in pCD201 | This study |
| pCpdBS261A | Replacement of Ser261 by Ala in CpdB in pCD201 | This study |
| pCpdBG242A | Replacement of Gly242 by Ala in CpdB in pCD201 | This study |
a
EcoRI restriction endonuclease site introduced by PCR design.