TABLE 1.

Strains, plasmids, and primers used in this study

Strain, plasmid, or oligonucleotideDescription or sequenceaSource or reference
E. coli
    DH5αhsdR recA lacZYA φ80lacZΔM15Promega
    JM109e14 mcrA recA1 endA1 gyrA96 thi-1 hsdR17(rK mK+) supE44 relA1 Δ(lac-proAB) [F′ traD36 proAB lacIqZΔM15]Promega
    MV118441
P. aeruginosa
    PAKNonmucoid P. aeruginosa strain KD. Bradley
    PAKpilA::TcrPreviously referred to as AWK44
    R306PAKpilV::Tn5-B212
    S39PAKalgR::Tn5-B2145
    PAKalgD::TcrTcr cassette insertion in EcoRI site of algDThis study
    PAKalgR7PAK encoding AlgR D54NThis study
    PAKalgR10PAK encoding AlgR D85NThis study
    PAKalgR11PAK encoding AlgR D54N D85NThis study
    PAO1Nonmucoid P. aeruginosa strain O1
    WFPA1algD::tet in PAO1This study
    WFPA12algR::aac1 in PAO1This study
    WFPA8algR7 in PAO1 (encoding AlgR D54N)This study
    WFPA13algR10 in PAO1 (encoding AlgR D85N)This study
    WFPA16algR11 in PAO1 (encoding AlgR D54N D85N)This study
    WFPA6algR gene replacement of WFPA12This study
Plasmids
    pSM-TETSource of Tcr cassette27
    pRIC380P. aeruginosa suicide vector1
    pUK21Kmr cloning vector42
    pUCPKSP. aeruginosa-E. coli shuttle vector43
    pGEMT-easyE. coli cloning vectorPromega
    pUCPAlgRalgR cloned into pUCPKS45
    pGemAlgRD54NalgR7 cloned into pGEMT-easyThis study
    pUCPAlgRD54NalgR7 cloned into pUCPKSThis study
    pDJW106algR cloned in pALTER-121
    pJK223R1Source of AlgR19
    pUS152algR7 in pALTER-121
    pUS164algR10 in pALTER-121
    pUS165algR11 in pALTER-121
    pUS162algR7 in pJK223R1 (source of AlgR D54N)This study
    pUS170algR10 in pJK223R1 (source of AlgR D85N)This study
    pUS172algR11 in pJK223R1 (source of AlgR D54N D85N)This study
    pUS150Wild-type algR in gene replacement vector pEX100T21
    pUS157algR7 in gene replacement vector pEX100T21
    pUS166algR10 in gene replacement vector pDJW52521
    pUS168algR11 in gene replacement vector pDJW52521
    pRS1Source of His-CheAR. Bourret
    pDJW220algD cloned in pALTER-15
Primers
    algR-Forward5′-TAAAGCGAGTCTCAGCGTCG-3′This study
    algR-Reverse5′-GGGACGACATGGGATATTCC-3′This study
    algD-Forward5′-CGCGGATCCGAGGTGAATGCGATGCG-3′This study
    algD-Reverse5′-TGCTCTAGACTAGGAGCAGATGCCCTC-3′This study
    algD335′-AGCCCTTGTGGCGAATAGGC-3′This study
    algD365′-GAATTGGGGAAAAGTCTGTG-3′This study
  • a Tcr, tetracycline resistance; aac1, gentamicin resistance; Kmr, kanamycin resistance.