Table 1.

Strains, phage, and plasmids used in this study

Strain, phage, or plasmidRelevant characteristicsSource or reference
Strains
E. coli
  XL1-Blue recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F′ proAB lacI q ZΔM15 Tn10(Tetr)]c Stratagene
  S17-1RP4 2-Tc::Mu-Km::Tn7 pro res mod + (TprSmr) 51
P. aeruginosa
  PA103Cytotoxic respiratory clinical isolate, Fla 38
  PA103ΔpilA PA103 withStuI/Bsu36I region of pilA replaced with a Gmr cassetteThis work
  PA103exoU::Tn5 Loss-of-cytotoxicity mutant 8; Tn5 Gmr inserted intoexoU 34
  PA103pscJ::Tn5 Loss-of-cytotoxicity mutant N; Tn5 Gmr inserted into pscJ; type III secretion defective, internalization competent 34
  PA103ΔpilApscJ::Tn5 PA103pscJ::Tn5with StuI/Bsu36I region of pilAreplaced with a Gmr cassetteThis work
Bacteriophage
 PO4 P. aeruginosabacteriophage 7
Plasmids
 pEX100TAllelic replacement suicide plasmid; Apr sacB oriT 49
 pX1918GTPlasmid containing axylE/aacC1 insertional cassette; Apr 49
 pP103pUC18 containing PA103 pilA 33
 pLW01pEX100T containing a 1.2-kb HindIII fragment of pilA with the StuI/Bsu36I region replaced with the xylE/aacC1 cassette of pX1918GTThis work
 pRIC276Tetr cassette;luxAB-containing plasmid55
 pLW02pLW01 with the XhoI/ClaI fragment of the aacC1 gene replaced with the Tetrcassette of pRIC276This work