TABLE 1.

Bacterial strains and plasmids

Strain or plasmidDescriptionaSource or reference
P. aeruginosa strains
    K767PAO1 wild typeN. Gotoh, Kyoto, Japan
    K1120K767 aphA22
    K1203K1120 ΔpvdDThis study
    K2333K1203 ΔfpvAThis study
    K2334K2333 attB::fpvA (WT)This study
    K2335K2333 attB::fpvA (W362A)This study
    K2340K2333 attB::fpvA (F366Y)bThis study
    K2341K2333 attB::fpvA (F369A)This study
    K2336K2333 attB::fpvA (W391A)This study
    K2343K2333 attB::fpvA (F795A)This study
    K2344K2333 attB::fpvA (Y796A)This study
    K2345K2333 attB::fpvA (Y801A)This study
Plasmidsd
    pSUP202ΔpvdDpSUP202::ΔpvdD; Tcr Apr Cmr Tra+I. L. Lamont, University of Otago, New Zealand
    mini-CTX1P. aeruginosa chromosome integration vector; Tcr11
    pCBS1mini-CTX1 ΔBamHI-SacIcThis study
    pCBS2pCBS1::fpvA (WT)This study
    pFLP2Carries gene for Flp recombinase; Apr Cbr11
  • a Amino acid changes in mutant FpvA proteins encoded by fpvA genes inserted into P. aeruginosa strain K2333 at the attB site are indicated in parentheses. WT, wild type.

  • b Mutation obtained via random mutagenesis of fpvA.

  • c mini-CTX1 derivative in which the BamHI and SacI sites have been engineered out of the vector.

  • d Plasmids mini-CTX1 and pCBS2 and their derivatives were maintained in E. coli and P. aeruginosa by the addition of tetracycline to the growth medium at 10 and 100 μg/ml, respectively. Plasmid pFLP2 was maintained in E. coli by the addition of ampicillin (100 μg/ml) and selected in P. aeruginosa by using carbenicillin (400 μg/ml). Plasmid pSUP202::ΔpvdD was maintained in E. coli by using 10 μg/ml tetracycline.