Table 3.
Strains and plasmids used in this study
| Characteristicsa | Source | |
|---|---|---|
| Strains | ||
| Vibrio fischeri | ||
| ES114 | wild-type V. fischeri | (Boettcher & Ruby, 1990) |
| AKD711 | ΔnsrR (allele exchanged from pAKD711 into ES114) |
this study |
| AKD712 | ΔnsrR, aox-lacZ chromosomal transcriptional fusion (exchanged from pAKD712 into AKD711) |
this study |
| AKD780 | Δaox (allele exchanged from pAKD780 into ES114) |
this study |
| AKD781 |
aox-lacZ chromosomal transcriptional fusion reporter (allele exchanged from pAKD712 into ES114) |
this study |
| AKD782 | ΔcydAB (allele exchanged from pAKD782 into ES114) |
this study |
| AKD783 | ΔcydAB, aox-lacZ chromosomal transcriptional fusion (exchanged from pAKD712 into AKD782) |
this study |
| AKD784 | Δaox ΔcydAB (allele exchanged from pAKD782 into AKD780) |
this study |
| AKD785 | ΔnsrR with aox-lacZ chromosomal transcriptional fusion reporter (allele exchanged from pAKD712 into AKD711) |
this study |
| AKD786 | Δaox ΔnsrR (allele exchanged from pAKD711 into AKD780) |
this study |
| AKD787 | ΔccoNOQP (allele exchanged from pAKD787 into ES114) |
this study |
| AKD788 | Δaox ΔccoNOQP (allele exchanged from pAKD787 into AKD780) |
this study |
| AKD789 | Suppressor mutant OG1-10 ΔccoNOQP (allele exchanged from pAKD787) | this study |
| Escherichia coli | ||
| DH5α | F’/endA1 hsdR17 glnV44 thi-1 recA1 gyrA relA1 Δ(lacIZYA-argF)U169 deoR (φ80dlacIΔ(lacZ)M15) |
(Hanahan, 1983) |
| DH5αλpir | λpir derivative of DH5α | (Dunn et al., 2005) |
| SASX41B | Hfr(PO2A), hemA41, relA1, spoT1, metB1 rrnB-2, mcrB1, creC510 |
E. coli genetic stock center |
| Plasmids | ||
| pVSV103 |
E. coli-V. fischeri shuttle vector, RP4 oriT pES213 and R6Kγ replication origins, lacZ, kanR |
(Dunn et al., 2006) |
| pVSV104 |
E. coli-V. fischeri shuttle vector, RP4 oriT pES213 and R6Kγ replication origins, lacZα, kanR |
(Dunn et al., 2006) |
| pVSV209 |
E. coli-V. fischeri shuttle vector, RP4 oriT pES213 and R6Kγ replication origins, promoterless gfp and chmR, constitutive rfp, kanR |
(Dunn et al., 2006) |
| pAKD601B | PCR-constructed derivative of pAKD600 this study (Dunn & Stabb, 2008) lacking gfp, contains lacIq and an IPTG-inducible promoter for overexpression of proteins in V. fischeri, kanR |
this study |
| pAKD601Baox | pAKD601B with the aox coding region | this study |
| pAKD602 | pVSV104 containing the aox coding region and the upstream intergenic region (Figure 1). |
this study |
| pAKD700 | pVSV209 digested with StuI and SanDI to remove gfp and chmR which was replaced with a StuI/SanDI lacZ fragment (lacZ PCR-amplified from pVSV103) |
this study |
| pAKD711 | ΔnsrR allele, R6Kγ and ColE1 replication origins, RP4 oriT, ermR, kanR |
this study |
| pAKD712 | aox-lacZ transcriptional fusion allele, R6Kγ and ColE1 replication origins, RP4 oriT, ermR, kanR |
this study |
| pAKD750 | pAKD700 with a 200 bp aox promoter region | this study |
| pAKD751 | pAKD750 PCR-amplified to introduce NsrR binding site modifications |
this study |
| pAKD780 | Δaox allele, R6Kγ and ColE1 replication origins, RP4 oriT, ermR, kanR |
this study |
| pAKD782 | ΔcydABybgTE allele, R6Kγ and ColE1 replication origins, RP4 oriT, ermR, kanR | this study |
| pAKD787 | ΔccoNOQP allele, R6Kγ and ColE1 replication origins, RP4 oriT, ermR, kanR |
this study |
| pEVS104 | conjugative helper plasmid, R6Kγ replication origin, kanR |
(Stabb & Ruby, 2002) |
a
abbreviations used: chmR, chloramphenicol resistance (cat); kanR, kanamycin resistance (aph); ermR, erythromycin resistance; rfp = red fluorescent protein