Table 1.

Bacterial strains and plasmids

Strain or plasmidGenotype or phenotypeReference or source
Strains
P. fluorescens
  CHA0Wild type 52
  CHA5 hcnB::Ω-Hg 53
  CHA21 anr::Ω-KmThis study (Fig.4)
  CHA77ΔhcnABC This study (Fig. 1)
  P3Wild type 53
P. aeruginosa
  PAO1Wild typeATCC 15692
  ADD1976Chromosomal insertion of T7 pol lacI q 2
E. coli
  BL21(DE3) hsdS gal; chromosomal insertion of T7 pol 44
  DH5α recA1 endA1 hsdR17 supE4 gyrA96 relA1Δ(lacZYA-argF)U169(φ80dlacZΔM15) 40
  JRG1728Δ(lacZYA)X74 galU galKΔ(ara-leu) rpsLΔ(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
 pEB16Broad-host-range vector; pBR322-pRO1600 replicon; RK2 Mob; Apr Cbr 2
 pHP45Ω-KmColE1 replicon; AprKmr 14
 pMMB67EHRSF1010 replicon; Apr 16
 pNM482ColE1 replicon; AprlacZ 34
 pVK100Broad-host-range vector; IncP replicon; RK2 Mob; TcrKmr 25
 pME497Mobilizing plasmid; IncP replicon; Tra; Apr 54
 pME3013pVK100 with an 8-kb HindIII genomic fragment of P. fluorescens CHA0 containing thehcnABC genes for HCN biosynthesis 53
 pME3071pMMB67 with a 5-kbXhoI-HindIII genomic fragment of CHA0 containing hcnABC′ This study (Fig. 1)
 pME3087Suicide vector; ColE1 replicon; RK2 Mob; Tcr 52
 pME3205pVK100 with a 3.8-kb SalI-HindIII fragment of pME3071 containing the hcnABC′ genes behind the kanamycin promoterThis study (Fig. 1)
 pME3206pVK100 with a 3.6-kbMaeI-HindIII fragment of pME3071 containing the hcnABC′ genes behind the kanamycin promoterThis study (Fig. 1)
 pME3209pEB16 with a 1.3-kbBamHI-HindIII fragment of pME3071 containing the hcnC′ gene behind the T7 promoterThis study (Fig. 1)
 pME3210pEB16 with a 3.6-kbMaeI-HindIII fragment of pME3071 containing the hcnABC′ genes behind the T7 promoterThis study (Fig.1)
 pME3212pEB16 with a 3.65-kbMaeI-StuI fragment of pME3071 containing thehcnABC genes behind the T7 promoterThis study (Fig. 1)
 pME3219pME6010 with a 410-bp hcnA promoter fragment and an hcnA′-′lacZ translational fusion at thePstI site in hcnA This study
 pME3580pKT240 with a 1.3-kb SacII genomic fragment of P. aeruginosa PAO1 containing the anrgene 17
 pME3812pVK100 with a 19-kbHindIII genomic fragment of P. fluorescensCHA0 containing the anr geneThis study
 pME3815pBluescript II KS+ with a 2.5-kbPstI fragment of pME3812 containing the anrgeneThis study (Fig. 4)
 pME3816pME3087 ΔEcoRIa with a 4.7-kb fragment containing anr::Ω-KmThis study (Fig. 4)
 pME3817pVK100 with a 1.6-kbBamHI-HindIII fragment of pME3818This study (Fig. 4)
 pME3818pME3815 with a deletion of the 0.9-kbEcoRV fragmentThis study (Fig. 4)
 pME6010pACYC177-pVS1 shuttle vector; Tcr S. Heeb (20a)
  • a ΔEcoRI signifies that theEcoRI site of pME3087 was deleted.