TABLE 1.
Strains and plasmids
| Strain or plasmid | Relevant characteristics | Reference |
|---|---|---|
| Strains | ||
| P. putida KT2442 | Rifr; derived from P. putida KT2440 | 15 |
| P. putida TEC1 | Rifr; P. putida KT2442 with a nearly complete internal deletion of pyrF | This work |
| P. putida TEC1 ΔbetCDE | Rifr Gmr; TEC1 whose betCDE putative operon was deleted by use of pTGLB | This work |
| E. coli CC118λpir | Δ(ara-leu) araD ΔlacX74 galE galK phoA20 thi-1 rpsE rpoB argE recA1 λpir lysogen | 15 |
| E. coli HB101 | Smr; rpsL recA thi pro leu hsdR−M+ (E. coli K-12/E. coli B. hybrid) | 37 |
| E. coli X82 | lacZ53(Am) λ−trpC50 pyrF287 hisG1(Fs) rpsL8; uracil auxotroph | E. coli Stock Center CGSC 5889 ID 6952 |
| Plasmids | ||
| pUC19NotI | Apr; pUC-type plasmid with MCS of pUC19 flanked by NotI sites | 15 |
| pUP | Apr; pUC19NotI inserted with a 2-kb EcoRI-HindIII PCR fragment containing the P. putida pyrF chromosome region | This work |
| pUPX | Apr; pUP with an xylE SmaI fragment from pXYLE10 cloned into the Eco47III and SmaI sites of pUP | This work |
| pKNG101 | Smr; sacRB+, broad-host-range plasmid of conditional replication | 17 |
| pKPX | Smr; pKNG1 with the pUPX ΔpyrF::xyIE NotI fragment | This work |
| pKT231 | Kmr, Smr, broad-host-range plasmid derived from RSF1010 | 10 |
| pKP | Kmr; pKT231 with the P. putida pyrF gene and its promoter region | This work |
| pGAPN | Apr; pGEM-T Easy carrying a PCR fragment of pyrF (with endogenous promoter) | This work |
| pGP704 | Apr; oriR6K, mobRK2, MCS of M13 tg131 | 25 |
| pGEM-T Easy | Apr; PCR fragment cloning plasmid | Promega |
| pGGLB | Apr; pGEM-T Easy with the betCDE deletion cassette cloned into the NotI site | Galväo et al., unpublished |
| pTEC | Kmr FOAs Ura+; ligation of MCS-Km-MCS, oriR6K/origin of transfer mobRK2, and pyrF PCR fragments | This work |
| pTGLB | Kmr Gmr FOAs Ura+; pTEC with the betCDE deletion cassette cloned into the NotI site | This work |
| pRK600 | Cmr; oriColE1, mobRK2, traRK2 | 19 |