TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Genotype or description | Reference or source |
|---|---|---|
| Strains | ||
| E. coli DH5α | F− φ80lacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK− mK+) phoA supE44 thi-1 gyrA96 relA1 λ− | Invitrogen |
| P. putida S12 | Wild type; srpABC+srpR+srpS+ | 5 |
| P. putida JK1 | srpB::TnMod-KmO | 4 |
| P. putida S12TS | srpS::ISPpu21 | This study |
| P. putida S12DM | srpS::ISPpu21 srpB::TnMod-KmO | This study |
| Plasmids | ||
| pCR2.1-TOPO | TOPO TA cloning vector for direct insertion of PCR products and blue/white screening; Apr Kmr | Invitrogen |
| pJD101 | Derived from BamHI digestion of P. putida JK1 chromosome; contains srpR through partial ORF of TnMod-KmO plasposon-mutated srpB | 9 |
| pJD102 | Derived from PstI digestion of P. putida JK1 chromosome; contains partial ORF of TnMod-KmO plasposon-mutated srpB through srpC | 9 |
| pJD203 | Constructed by digestion (with EcoRV and BamHI) and ligation of pJD101 and pJD102; contains partial ORF of TnMod-KmO plasposon-mutated srpB through srpC | This study |
| pBBR1MCS-2 | Broad-host-range cloning vector; Kmr | 11 |
| pBBR1-2-srpR | Coding region of srpR cloned into pBBR1MCS-2 (using EcoRI and BamHI sites) | This study |
| pBBR1-2-srpS | Coding region of srpS cloned into pBBR1MCS-2 (using EcoRI and BamHI sites) | This study |
| pBBR1-2-srpRS | Coding regions of srpR and srpS cloned into pBBR1MCS-2 (using EcoRI and BamHI sites) | This study |