TABLE 1.
Bacterial strains and plasmids used in this work
AmpR, ampicillin resistance; CmR, chloramphenicol resistance; GmR, gentamicin resistance; KanR, kanamycin resistance; TetR, tetracycline resistance.
| Relevant characteristics | Source | |
|---|---|---|
| Bacterial strain | ||
| JRG6348 | A derivative of E. coli JRG1728 lac, fnr carrying a chromosomal FNR-dependent promoter-lacZ fusion; CmR | Dr. David Lee (University of Birmingham, Birmingham, UK) |
| JRG6725 | P. putida KT2440 parent strain; source of genomic DNA for amplification and cloning anr, PP_3233, and PP_3287 genes for protein expression and construction of JRG6721, JRG6722, and JRG6723 | Laboratory collection |
| JRG6721 | P. putida KT2440 PP_3233, anr (PP_3287+) | This work |
| JRG6722 | P. putida KT2440 PP_3287, anr (PP_3233+) | This work |
| JRG6723 | P. putida KT2440 PP_3233, PP_3287 (ANR+) | This work |
| Plasmid | ||
| pAS12 | pEX18Ap containing gene replacement cassette for PP_4265, AmpR, GmR | This study |
| pBBR1-MCS-5 | Broad host range vector, KanR. | Ref. 35 |
| pDelta_PP_3233 | pEMG containing replacement cassette for PP3233, KanR | This study |
| pDelta_PP_3287 | pEMG containing replacement cassette for PP3287, KanR | This study |
| pEMG | Gene replacement vector with two flanking I-SceI sites, KanR | Ref. 23 |
| pEX18Ap | Gene replacement vector, sacB, AmpR | Ref. 22 |
| pFLP2 | Broad host range vector with FLP recombinase, AmpR | Ref. 22 |
| pGS422 | pUC13 containing the FF-41.5 promoter (EcoRI-HindIII) | Ref. 36 |
| pGS652 | pBluescript containing the NN-41.5 promoter (EcoRI-HindIII) | Ref. 37 |
| pGS810 (pFF-41.5) | pRW50 derivative with lacZ under the control of a class II FNR-dependent promoter, TetR | Ref. 38 |
| pGS2268 | pGEX-KG (39) containing ANR for expression as a GST fusion protein; AmpR | This work |
| pGS2350 | pBAD-HisB (Invitrogen) derivative for expression of E. coli fnr, AmpR | This work |
| pGS2351 | pBAD-HisB derivative for expression of P. putida anr; AmpR | This work |
| pGS2352 | pBAD-HisB derivative for expression of P. putida PP_3233, AmpR | This work |
| pGS2353 | pBAD-HisB derivative for expression of P. putida PP_3287, AmpR | This work |
| pGS2403 | As for pGS2414 but containing PP_3233 | This work |
| pGS2413 | As for pGS2414 but containing PP_3287 | This work |
| pGS2414 | pCOLD-TF (Takara Clontech) containing anr for expression as a His6-Tig fusion protein, AmpR | This work |
| pGS2508 | PP_3233 ligated into pBB1MSC-5 (EcoRI-XhoI) for expression of PP_3233 in P. putida, GmR | This work |
| pGS2509 | PP_3287 ligated into pBB1MSC-5 (EcoRI-XhoI) for expression of PP_3287 in P. putida; GmR | This work |
| pPS858 | Source of gentamicin resistance cassette; AmpR, GmR | Ref. 22 |
| pSWI | Broad host range vector with I-SceI endonuclease, AmpR | Ref. 40 |