TABLE 1.
Bacterial strains and plasmids used in this study
| Bacterial straina or plasmid | Relevant genotype or characteristicsb | Reference(s) or source |
|---|---|---|
| Bacterial strains | ||
| P. aeruginosa | ||
| WT | Wild-type P. aeruginosa PAO1; strain H103 | 3, 47 |
| UW-WT | UW wild-type P. aeruginosa PAO1 | 26 |
| UW-PA1799 | PA1799::ISlacZ/hah-Tcr; insertion at 492 (708) bp in PA1799; derived from strain UW-WT | 26 |
| UW-PA1798 | PA1798::ISlacZ/hah-Tcr; insertion at 307 (1,287) bp in PA1798; derived from UW-WT | 26 |
| parR mutant | PA1799::ISlacZ/hah-Tcr, H103 background; Tetr | This study |
| parS mutant | PA1798::ISlacZ/hah-Tcr, H103 background; Tetr | This study |
| parR (Tn7-parRS+) complemented strain | PA1799 mutant with Tn7-PA1798-1799+ integrated; Tcr Gmr | This study |
| UW-PA1797 | PA1797::ISlacZ/hah-Tcr; insertion at 423 (1,833) bp in PA1797; derived from UW-WT | 26 |
| UW-PA1559 | PA1559::ISlacZ/hah-Tcr; insertion at 406 (732) bp in PA1559; derived from UW-WT | 26 |
| UW-pagL | pagL::ISphoA/hah-Tcr; insertion at 196 (522) bp in pagL; derived from UW-WT | 26 |
| UW-PA4782 | PA4782::ISphoA/hah-Tcr; insertion at 147 (246) bp in PA4782; derived from UW-WT | 26 |
| UW-pmrB | pmrB::ISphoA/hah-Tcr; insertion at 67 (1,434) bp in pmrB; derived from UW-WT | 26 |
| UW-arnB | arnB::ISphoA/hah-Tcr; insertion at 327 (1,149) bp in arnB; derived from UW-WT | 26 |
| UW-arnC | arnC::ISlacZ/hah-Tcr; insertion at 408 (1,020) bp in arnC; derived from UW-WT | 26 |
| E. coli TOP10 | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZM15 ΔlacX74recA1araΔ139 Δ(ara-leu)7697galUgalKrpsL (Strr) endA1nupG | Invitrogen |
| Plasmids | ||
| pCR-Blunt II-TOPO | PCR cloning vector; Kanr | Invitrogen |
| pCR-PA1798-1799+ | pCR-Blunt II-TOPO harboring 2.3-kb PA1798-PA1799 amplicon | This study |
| pUC18-mini-Tn7T-Gm | Suicide plasmid; Gmr Ampr | 5 |
| pUC-Tn7-PA1798-1799+ | pUC18-mini-Tn7T-Gm with 2.3-kb PA1798-PA1799 fragment from pCR-PA1798-PA1799 | This study |
| pTNS2 | Transposition helper plasmid; Ampr | 5 |
| pUCPlux-pPA3552 | pUCP23 containing a transcriptional fusion between the arnB promoter and luxCDABE | 36 |
a
UW, University of Washington Genome Science Center.
b
Antibiotic resistance phenotypes: Ampr, ampicillin resistance for E. coli and carbenicillin resistance for P. aeruginosa; Gmr, gentamicin resistance; Kanr, kanamycin resistance; Tcr, tetracycline resistance. An “insertion at 492 (708) bp” means that the insertion is at nucleotide 492 within a 708-bp-long gene. “hah” is a 63-codon insertion which encodes an influenza hemagglutinin epitope together with a hexahistidine metal affinity purification tag.