TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Genotype or phenotype | Reference or source |
|---|---|---|
| Strains | ||
| P. fluorescens | ||
| CHA0 | Wild type | 52 |
| CHA5 | hcnB::Ω-Hg | 53 |
| CHA21 | anr::Ω-Km | This study (Fig. 4) |
| CHA77 | ΔhcnABC | This study (Fig. 1) |
| P3 | Wild type | 53 |
| P. aeruginosa | ||
| PAO1 | Wild type | ATCC 15692 |
| ADD1976 | Chromosomal insertion of T7 pol lacIq | 2 |
| E. coli | ||
| BL21(DE3) | hsdS gal; chromosomal insertion of T7 pol | 44 |
| DH5α | recA1 endA1 hsdR17 supE4 gyrA96 relA1 Δ(lacZYA-argF)U169 (φ80dlacZΔM15) | 40 |
| JRG1728 | Δ(lacZYA)X74 galU galK Δ(ara-leu) rpsL Δ(tyrR-fnr-trg) | 17; J. R. Guest |
| RU4420 | thi-1 endA1 hsdR17 supE44 trp::Tn1725 | 47 |
| Plasmids | ||
| pBluescript II KS+ | Cloning vector; ColE1 replicon; Apr | Stratagene |
| pEB16 | Broad-host-range vector; pBR322-pRO1600 replicon; RK2 Mob; Apr Cbr | 2 |
| pHP45Ω-Km | ColE1 replicon; Apr Kmr | 14 |
| pMMB67EH | RSF1010 replicon; Apr | 16 |
| pNM482 | ColE1 replicon; Apr ′lacZ | 34 |
| pVK100 | Broad-host-range vector; IncP replicon; RK2 Mob; Tcr Kmr | 25 |
| pME497 | Mobilizing plasmid; IncP replicon; Tra; Apr | 54 |
| pME3013 | pVK100 with an 8-kb HindIII genomic fragment of P. fluorescens CHA0 containing the hcnABC genes for HCN biosynthesis | 53 |
| pME3071 | pMMB67 with a 5-kb XhoI-HindIII genomic fragment of CHA0 containing hcnABC′ | This study (Fig. 1) |
| pME3087 | Suicide vector; ColE1 replicon; RK2 Mob; Tcr | 52 |
| pME3205 | pVK100 with a 3.8-kb SalI-HindIII fragment of pME3071 containing the hcnABC′ genes behind the kanamycin promoter | This study (Fig. 1) |
| pME3206 | pVK100 with a 3.6-kb MaeI-HindIII fragment of pME3071 containing the hcnABC′ genes behind the kanamycin promoter | This study (Fig. 1) |
| pME3209 | pEB16 with a 1.3-kb BamHI-HindIII fragment of pME3071 containing the hcnC′ gene behind the T7 promoter | This study (Fig. 1) |
| pME3210 | pEB16 with a 3.6-kb MaeI-HindIII fragment of pME3071 containing the hcnABC′ genes behind the T7 promoter | This study (Fig. 1) |
| pME3212 | pEB16 with a 3.65-kb MaeI-StuI fragment of pME3071 containing the hcnABC genes behind the T7 promoter | This study (Fig. 1) |
| pME3219 | pME6010 with a 410-bp hcnA promoter fragment and an hcnA′-′lacZ translational fusion at the PstI site in hcnA | This study |
| pME3580 | pKT240 with a 1.3-kb SacII genomic fragment of P. aeruginosa PAO1 containing the anr gene | 17 |
| pME3812 | pVK100 with a 19-kb HindIII genomic fragment of P. fluorescens CHA0 containing the anr gene | This study |
| pME3815 | pBluescript II KS+ with a 2.5-kb PstI fragment of pME3812 containing the anr gene | This study (Fig. 4) |
| pME3816 | pME3087 ΔEcoRIa with a 4.7-kb fragment containing anr::Ω-Km | This study (Fig. 4) |
| pME3817 | pVK100 with a 1.6-kb BamHI-HindIII fragment of pME3818 | This study (Fig. 4) |
| pME3818 | pME3815 with a deletion of the 0.9-kb EcoRV fragment | This study (Fig. 4) |
| pME6010 | pACYC177-pVS1 shuttle vector; Tcr | S. Heeb (20a) |
a
ΔEcoRI signifies that the EcoRI site of pME3087 was deleted.