TABLE 1.

Bacterial strains and plasmids used in this study

Bacterial straina or plasmidRelevant genotype or characteristicsbReference(s) or source
Bacterial strains
    P. aeruginosa
        WTWild-type P. aeruginosa PAO1; strain H1033, 47
        UW-WTUW wild-type P. aeruginosa PAO126
        UW-PA1799PA1799::ISlacZ/hah-Tcr; insertion at 492 (708) bp in PA1799; derived from strain UW-WT26
        UW-PA1798PA1798::ISlacZ/hah-Tcr; insertion at 307 (1,287) bp in PA1798; derived from UW-WT26
        parR mutantPA1799::ISlacZ/hah-Tcr, H103 background; TetrThis study
        parS mutantPA1798::ISlacZ/hah-Tcr, H103 background; TetrThis study
        parR (Tn7-parRS+) complemented strainPA1799 mutant with Tn7-PA1798-1799+ integrated; Tcr GmrThis study
        UW-PA1797PA1797::ISlacZ/hah-Tcr; insertion at 423 (1,833) bp in PA1797; derived from UW-WT26
        UW-PA1559PA1559::ISlacZ/hah-Tcr; insertion at 406 (732) bp in PA1559; derived from UW-WT26
        UW-pagLpagL::ISphoA/hah-Tcr; insertion at 196 (522) bp in pagL; derived from UW-WT26
        UW-PA4782PA4782::ISphoA/hah-Tcr; insertion at 147 (246) bp in PA4782; derived from UW-WT26
        UW-pmrBpmrB::ISphoA/hah-Tcr; insertion at 67 (1,434) bp in pmrB; derived from UW-WT26
        UW-arnBarnB::ISphoA/hah-Tcr; insertion at 327 (1,149) bp in arnB; derived from UW-WT26
        UW-arnCarnC::ISlacZ/hah-Tcr; insertion at 408 (1,020) bp in arnC; derived from UW-WT26
    E. coli TOP10F mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZM15 ΔlacX74 recA1 araΔ139 Δ(ara-leu)7697 galU galK rpsL (Strr) endA1 nupGInvitrogen
Plasmids
    pCR-Blunt II-TOPOPCR cloning vector; KanrInvitrogen
    pCR-PA1798-1799+pCR-Blunt II-TOPO harboring 2.3-kb PA1798-PA1799 ampliconThis study
    pUC18-mini-Tn7T-GmSuicide plasmid; Gmr Ampr5
    pUC-Tn7-PA1798-1799+pUC18-mini-Tn7T-Gm with 2.3-kb PA1798-PA1799 fragment from pCR-PA1798-PA1799This study
    pTNS2Transposition helper plasmid; Ampr5
    pUCPlux-pPA3552pUCP23 containing a transcriptional fusion between the arnB promoter and luxCDABE36
  • 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.