TABLE 1.
P. aeruginosa strains and plasmids used in this study
| Strain or plasmid | Antibiotic resistancea | Description | Source, reference, or accession no. |
|---|---|---|---|
| E. coli SM10 | None | Host strain for plasmids pEXAp18, pMMB67, mini-CTX, and mini-CTX-lacZ and their derivatives | Lab collection |
| P. aeruginosa strains | |||
| PAO1 | None | Laboratory strain | Lab collection |
| PAO1pilL2P | None | Strain PAO1 with the DNA fragment containing putative promoter of the PAPI-1 pil operon and a promoterless lacZ gene inserted at the CTX phage att site on the chromosome; the backbone of the plasmid mini-CTX-lacZ was eliminated | This study |
| PAO1pilS2P | None | Strain PAO1 with a DNA fragment containing the putative promoter of the pilS2 gene and a promoterless lacZ gene inserted at the CTX phage att site on the chromosome; the backbone of the plasmid mini-CTX-lacZ was eliminated | This study |
| PAO1Bla6 | Cbr | Strain PAO1 with genes bla and lacZ inserted at the CTX phage att site on the chromosome | 27 |
| PA14 | None | Burn isolate | 27 |
| PA14pilL2P | None | Strain PA14 with a DNA fragment containing the putative promoter of the PAPI-1 pil operon (upstream of the pilL2 gene) and a promoterless lacZ gene inserted at the CTX phage att site on the chromosome; the backbone of the plasmid mini-CTX-lacZ was eliminated | This study |
| PA14pilS2P | None | Strain PA14 with a DNA fragment containing the putative promoter of the pilS2 gene and a promoterless lacZ gene inserted at the CTX phage att site on the chromosome; the backbone of the plasmid mini-CTX-lacZ was eliminated | This study |
| PA14 ΔpilS2ppilS2-His | Tcr | Deletion mutant PA14 ΔpilS2 carrying the cloned piS2-His in plasmid pMMB67Tc (ppilS2-His) | This study |
| PA14 ΔpilDΔpilS2ppilS2-His | Tcr | Deletion mutant PA14 ΔpilD ΔpilS2 carrying the cloned piS2-His in plasmid pMMB67Tc (ppilS2-His) | This study |
| PA14 ΔpilDatt::pilDΔpilS2ppilS2-His | Tcr | Strain PA14 ΔpilDΔpilS2ppilS2-His with the pilD gene driven by Ptac inserted at the att site on the chromosome; the backbone of the mini-CTX was eliminated | This study |
| PA14TnC2 | Gmr | Strain PA14 with a transposon MAR2×T7 inserted at nucleotide 1634 of PAPI-1 gene RL090 (PA14_59200) | 22 |
| PA14TnC2 ΔpilS2 | Gmr | In-frame deletion of the PAPI-1 pilS2 gene in strain PA14TnC2 | This study |
| PA14TnC2 ΔpilV2 | Gmr | In-frame deletion of the PAPI-1 pilV2 gene in strain PA14TnC2 | This study |
| PA14TnC2 ΔpilA | Gmr | In-frame deletion of the pilA gene in strain PA14TnC2 | This study |
| PA14TnC2 Δflp | Gmr | In-frame deletion of the flp gene in strain PA14TnC2 | This study |
| PA14TnC2 ΔfppA | Gmr | In-frame deletion of the fppA gene in strain PA14TnC2 | This study |
| PA14TnC2 ΔpilD | Gmr | In-frame deletion of the pilD gene in strain PA14TnC2 | This study |
| PA14TnC2 ΔpilAΔpilS2 | Gmr | In-frame deletion of both the pilA and pilS2 genes in strain PA14TnC2 | This study |
| PA14TnC2 ΔpilS2ppilS2 | Gmr Tcr | Strain PA14TnC2 ΔpilS2 carrying cloned pilS2 on plasmid pMMB67Tc (ppilS2) | This study |
| PA14TnC2 ΔpilV2ppilV2 | Gmr Tcr | Strain PA14TnC2 ΔpilV2 carrying cloned pilV2 on plasmid pMMB67Tc (ppilV2) | This study |
| PA14TnC2 ΔpilS2vector | Gmr Tcr | Strain PA14TnC2 ΔpilS2 carrying plasmid pMMB67Tc | This study |
| PA14TnC2 ΔpilV2vector | Gmr Tcr | Strain PA14TnC2 ΔpilV2 carrying plasmid pMMB67Tc | This study |
| PA14TnC2 ΔpilDppilD | Gmr Tcr | Strain PA14TnC2 ΔpilD carrying the cloned pilD gene on plasmid pMMB67Tc (ppilD) | This study |
| PA14TnC2 ΔpilDpfppA | Gmr Tcr | Strain PA14TnC2 ΔpilD carrying the cloned fppA gene on plasmid pMMB67Tc (pfppA) | This study |
| PA14TnC2 ΔpilDvector | Gmr Tcr | Strain PA14TnC2 ΔpilD carrying plasmid pMMB67Tc | This study |
| PA14TnC2 ΔpilAΔpilS2vector | Gmr Tcr | Strain PA14TnC2 ΔpilAΔpilS2 carrying plasmid pMMB67Tc | This study |
| PA14TnC2 ΔpilAΔpilS2ppilS2 | Gmr Tcr | Strain PA14TnC2 ΔpilAΔpilS2 carrying plasmid ppilS2 | This study |
| PA14TnC2 ΔpilAΔpilS2ppilS2-Flag | Gmr Tcr | Strain PA14TnC2 ΔpilAΔpilS2 carrying plasmid ppilS2-Flag, in which the Flag tag is fused with PilS2 at its C terminus | This study |
| Plasmids | |||
| pEXG2 | Gmr | Gene replacement vector for constructing deletion or insertion mutants of P. aeruginosa | 28 |
| pEX18Ap | Apr | Gene replacement vector for constructing deletion or insertion mutants of P. aeruginosa | 12 |
| pFLP2 | Apr | Plasmid used for resolving the backbone of mini-CTX-lacZ | 12 |
| Mini-CTX | Tcr | Gene delivery vector for inserting genes at the CTX phage att site on the P. aeruginosa chromosome | 13 |
| CTX-PtacpilD | Tcr | The pilD gene and Ptac were cloned in the vector mini-CTX | This study |
| Mini-CTX-lacZ | Tcr | Gene delivery vector for inserting genes at the CTX phage att site on the P. aeruginosa chromosome | AF140579 |
| CTXpilL2P | Tcr | A DNA fragment containing the putative promoter of the PAPI-1 pil operon was cloned in the vector mini-CTX-lacZ | This study |
| CTXpilS2P | Tcr | A DNA fragment containing the putative promoter of the PAPI-1 pilS2 gene was cloned in the vector mini-CTX-lacZ | This study |
| pMMB67Tc | Tcr | Replicative plasmid in P. aeruginosa, used for complementary analysis in P. aeruginosa deletion mutants | 40 |
| ppilS2 | Tcr | The pilS2 gene was cloned in plasmid pMMB67Tc | This study |
| ppilV2 | Tcr | The pilV2 gene was cloned in plasmid pMMB67Tc | This study |
| ppilD | Tcr | The pilD gene was cloned in plasmid pMMB67Tc | This study |
| ppilS2-His | Tcr | The pilS2 gene and His tag at the C terminus were cloned in plasmid pMMB67Tc | This study |
| ppilS2-Flag | Tcr | The pilS2 gene and Flag tag at the C terminus were cloned in plasmid pMMB67Tc | This study |
a
Apr, ampicillin resistance; Gmr, gentamicin resistance; Cbr, carbenicillin resistance; Tcr, tetracycline resistance.