TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Genotype or construction | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α | supE44 ΔlacU169 (φ80dlacZΔM15) recA1 endA1 hsdR17 thi-1 gyrA96 relA1 | Invitrogen |
| HB101 | subE44 subF58 hsdS3(rB− mB−) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 mtl-1 | 10 |
| CC118 λpir | Δ(ara-leu) araD ΔlacX74 galE galK phoA20 thi-1 rpsE rpoB argE(Am) recA1 λpir phage lysogen | 28 |
| P. putida | ||
| PaW85 | WT | 7 |
| PaWRpoS | PaW85, rpoS::km (the same as PKS54) | 55 |
| PaWRpoSLacI | PaWRpoS, lacIq gene in chromosome | This study |
| PaWRpoSSodAB | PaWRpoS, lacIq-Ptac-sodAB expression cassette in chromosome | This study |
| PaWRpoSKatA | PaWRpoS, Ptac-katA transcriptional fusion and lacIq gene in chromosome | This study |
| PaWRpoSSodABKatA | PaWRpoS, lacIq-Ptac-sodAB and Ptac-katA expression cassettes in chromosome | This study |
| Plasmids | ||
| pBluescript KS(+) | Cloning vector (Apr) | Stratagene |
| pUC18Not | Cloning vector (Apr) | 28 |
| pUTmini-Tn5 Cm | Delivery plasmid for mini-Tn5 Cm (Apr Cmr) | 65 |
| pJMT6 | Delivery plasmid for mini-Tn5 Tel (Apr Telr) | 65 |
| pRK2013 | Helper plasmid for conjugal transfer of mini-Tn-carrying plasmids (Kmr) | 18 |
| pBK-miniTn7-ΩSm1 | pUC19-based delivery plasmid for mini-Tn7-ΩSm1 (Smr Apr) | 34 |
| pUX-BF13 | R6K replicon-based helper plasmid providing Tn7 transposase proteins (Apr Mob+) | 6 |
| pAM10.6 | Biodegradative plasmid carrying catalase gene katA | 57 |
| pBRlacItac | pBR322 carrying Ptac promoter and lacIq gene for LacI repressor | 55 |
| pKSsodB | pBluescript KS (+) containing PCR-amplified sodB gene region inserted into EcoRV-opened vector plasmid | This study |
| pKSsodAB | PCR-amplified sodA gene in SmaI site of pKSsodB | This study |
| pBRlacItacsodAB | pBRlacItac with sodAB genes cloned from pKSsodAB within XbaI- and SalI-generated DNA fragment | This study |
| pUC18NotlacItacsodAB | pUC18Not containing lacIq-Ptac-sodAB expression cassette cloned within BamHI-generated DNA fragment from pBRlacItacsodAB | This study |
| pUTtellacItacsodAB | mini-Tn5 delivery plasmid pJMT6 containing lacIq-Ptac-sodAB expression cassette cloned from pUC18NotlacItacsodAB in NotI site | This study |
| pACtac | pACYC184 containing Ptac promoter | 71 |
| pKStac | pBluescript KS(+) containing Ptac promoter cloned from plasmid pACtac within BamHI- and HindIII-generated DNA fragment | This study |
| pKStackatA | PCR-amplified katA gene from plasmid pAM10.6 inserted into XbaI- and NotI-cleaved pKStac | This study |
| pAKNtackatA | pBK-miniTn7-ΩSm1 containing Ptac-katA transcriptional fusion inserted into ApaI- and NotI-cleaved vector plasmid | This study |
| pREP4 | LacI repressor-expressing plasmid | Qiagen |
| pUC18NotlacI | pUC18Not containing lacIq gene from pREP4 cloned within Eco47III- and SmaI-generated fragment into SmaI site | This study |
| pUTCmlacI | lacIq gene from pUC18NotlacI inserted within NotI fragment into mini-Tn5 delivery plasmid pUTmini-Tn5 Cm | This study |
| pKTpheA56+A | Test system for detection of Phe+ revertants occurring due to 1-bp deletions | 74 |
| pKTpheA22TAG | Test system for detection of Phe+ revertants occurring due to base substitutions | 74 |