TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Genotype or phenotype | Reference or source |
|---|---|---|
| Strains | ||
| P. aeruginosa | ||
| PAO1 | Prototroph, chl-2 | 23 |
| ADD1976 | Prototroph, chl-2 Tcr | 8 |
| ADD1976ΔtatC | ΔtatC::Gm | This study |
| ADD1976ΔtatC PlcH | ΔtatC::Gm::plcH | This study |
| ADD1976ΔtatC PlcH R34A | ΔtatC::Gm::plcHR34A | This study |
| ADD1976ΔtatC PlcH R35A | ΔtatC::Gm::plcHR35A | This study |
| ADD1976ΔtatC PlcH R34AR35A | ΔtatC::Gm::plcHR34AR35A | This study |
| PAO1 ΔHRN | ΔplcH ΔplcN ΔplcR1 ΔplcR2 | 44 |
| PAO1ΔHRN PlcH | ΔplcH ΔplcN ΔplcR1 ΔplcR2::plcH | This study |
| PAO1ΔHRN PlcH R9K | ΔplcH ΔplcN ΔplcR1 ΔplcR2::plcHR9K | This study |
| PAO1ΔHRN PlcH R10K | ΔplcH ΔplcN ΔplcR1 ΔplcR2::plcHR10K | This study |
| PAO1ΔtatC | ΔtatC::Gm | 33 |
| PAO1Δtat operon | ΔtatA ΔtatB ΔtatC ΔPA5071 | This study |
| E. coli | ||
| BL21-DE3(pLysS) | hsdS gal ompT rB− mB− (cIts857 ind1 Sam7 nin5 lacUV5 T7) | 47 |
| Plasmids | ||
| pUCP19 | Cbr pUC derivative, P. aeruginosa origin of replication | 8 |
| pUC18plcH | plcHR1R2 3.2-kb BamHI-SphI | 46 |
| pADD3268 | Broad host range, Apr Cbr T7 promoter | 8 |
| pADDplcH | plcHR 3.2-kb BamHI/HindIII | 46 |
| pADDplcHR8K | PlcH R8K point mutation | This study |
| pADDplcHR8S | PlcH R8S point mutation | This study |
| pADDplcHR8A | PlcH R8A point mutation | This study |
| pADDplcHR9K | PlcH R9K point mutation | 33 |
| pADDplcHR10K | PlcH R10K point mutation | This study |
| pADDplcHR34A | PlcH R34A point mutation | This study |
| pADDplcHR35A | PlcH R35A point mutation | This study |
| pADDplcHR34R35A34A35 | PlcH R34AR35A double mutation | This study |
| pADDplcHΔss | PlcH Δaa 1-35 signal sequence mutation | This study |
| pUCP19E.c tat | Cbr pUC derivative, Pseudomonas origin of replication 2-kb SpeI E. coli tatABC | This study |
| pUCP19P.a tat | Cbr pUC derivative, Pseudomonas origin of replication 4-kb PstI P. aeruginosa tatABC | This study |