Table 1.
Strains and plasmids used in this study
| Strain/plasmid | Relevant characteristics | Source |
|---|---|---|
| Strains | ||
| Escherichia coli | ||
| DH5α | General purpose cloning strain; Δ(lacZ)M15 | New England Biolabs |
| DBS206 | E. coli DH5α harboring 3x-V5 tag on pCR2.1 TOPO | This study |
| DBS215 | DH5α with ampR ORF and the ampR-ampC intergenic region PCR cloned into pCR2.1 TOPO | This study |
| DBS222 | DH5α harboring ampR ORF tagged with 3x V5-tag on pCR2.1 TOPO | This study |
| DBS234 | DH5α with mini CTX2, containing 3x V5-tagged ampR | This study |
| Pseudomonas aeruginosa | ||
| PAO1 | Wild-type | (23) |
| PKM315 | PAOΔampR; in-frame deletion of ampR (PA4109) | (21) |
| DBS248 | PAOΔampR::ampR-V5; 3x-V5-tagged ampR cloned onto mini-CTX2 and moved into PKM315 | This study |
| Plasmids | ||
| pCR2.1 TOPO | TA cloning vector for PCR products; ApR, KmR; ColE1 f1 ori lacZα | Invitrogen |
| ZM747-V5 | 3x V5 tag C-term and N-term of yeast URA-3 in pBlueScript; pDBS193 | (24) |
| Mini CTX2 | Plasmid for single-copy gene integration in P. aeruginosa; TcR | (25) |
| pDBS206 | 3x V5 tag PCR-amplified from ZM747-V5 and cloned into pCR2.1 TOPO | This study |
| pDBS215 | 1112-bp ampR ORF along with the ampR-ampC intergenic region, PCR-amplified with primers DBS_ampRF1 and DBSampR; cloned into pCR2.1 TOPO | This study |
| pDBS222 | ampR ORF from pDBS215 subcloned as KpnI-SacI upstream of and inframe with 3x V5 tag in pDBS206 | This study |
| pDBS234 | 3x V5-tagged ampR ORF from pDBS222 subcloned as a KpnI-NotI fragment into mini-CTX2 | This study |