TABLE 1.
Bacterial strains and plasmids used in this study
| Bacterial strain or plasmid | Relevant characteristics and/or description | Source or reference |
|---|---|---|
| E. coli strains | ||
| DH5α | φ80dlacZΔM15 endA1 recA1 gyrA96 thi-1 hsdR17 (rK− mK+) relA1 supE44 deoR Δ(lacZYA-argF)U196 | Promega Corp., Madison, WI |
| JM109 | endA1 gyrA96 hsdR17(rK− mK+) mcrB+recAl relA1 supE44 thi-1 Δ(lac-proAB) F′ [traD36 proAB lacIqZΔM15] | 26 |
| Plasmid vectors | ||
| pACYC177 | Apr Kmr; p15A replicon; cloning vector | 6 |
| pBAD28 | Apr Cmr; arabinose-inducible expression vector; pACYC184 replicon | 11 |
| pBR322 | Apr Tcr; ColE1 replicon; cloning vector | 3 |
| pOU82 | Apr; lacZYA; R1 replicon | 10 |
| Plasmid constructs | ||
| pACYC177ΔApr | Kmr Amps; 683-bp deletion of a BamHI-ScaI fragment from pACYC177 | This study |
| pACYC177-TA(Apr) | Apr; 731-bp PCR fragment containing pRAS3.1 pemIK-like genes (nucleotide positions 8819 to 8088) cloned into the XhoI-BamHI sites of pACYC177 | This study |
| pACYC177-TA(Kmr) | Kmr; 731-bp PCR fragment containing pRAS3.1 pemIK-like genes (nucleotide positions 8819 to 8088) cloned into the BamHI-ScaI sites of pACYC177 | This study |
| pBAD28-repC | Apr Cmr; 1,015-bp PCR fragment containing the pRAS3.1 repC (nucleotide positions 6903 to 5889) cloned behind the PBAD promoter | 17 |
| pOU82-TA | Apr; 731-bp PCR fragment containing pRAS3.1 pemIK-like genes (nucleotide positions 8819 to 8088) cloned into pOU82 | 17 |
| pRAS3.1 | Tcr; natural 11,851-bp plasmid isolated from Aeromonas salmonicida subsp. salmonicida with four iterons and five 6-bp repeats | 12 |
| pRAS3.1Km | Kmr; pRAS3.1 with Tcr replaced by Kmr from pSKm2 at the BamHI-EcoRV sites | 17 |
| pRAS3.1.34 | Tcr; pRAS3.1.35 derivative with four 6-bp repeats from pRAS3.2 by exchanging the 2.9-kb HindIII-PvuI region | This study |
| pRAS3.1.35 | Tcr; pRAS3.1 derivative with three iterons obtained by random ligation of short iteron fragments after BstEII digestion | This study |
| pRAS3.1.35Km | Kmr; pRAS3.1.35 with Tcr replaced by Kmr from pSKm2 at the BamHI-EcoRV sites | This study |
| pRAS3.1.44 | Tcr; pRAS3.1 derivative with four 6-bp repeats from pRAS3.2 by exchanging the 2.9-kb HindIII-PvuI region | This study |
| pRAS3.1.55 | Tcr; pRAS3.1 derivative with five iterons obtained by random ligation of short iteron fragments after BstEII digestion | This study |
| pRAS3.1.55Km | Kmr; pRAS3.1.55 with Tcr replaced by Kmr from pSKm2 at the BamHI-EcoRV sites | This study |
| pRAS3.1.74 | Tcr; pRAS3.1.75 derivative with four 6-bp repeats from pRAS3.2 by exchanging the 2.9-kb HindIII-PvuI region | This study |
| pRAS3.1.75 | Tcr; pRAS3.1 derivative with seven iterons obtained by random ligation of short iteron fragments after BstEII digestion | This study |
| pRAS3.1.75Km | Kmr; pRAS3.1.75 with Tcr replaced by Kmr from pSKm2 at the BamHI-EcoRV sites | This study |
| pRAS3.2 | Tcr; natural 11,823-bp plasmid isolated from atypical Aeromonas salmonicida with three iterons and four 6-bp repeats | 12 |
| pRAS3.2Km | Kmr; pRAS3.2 with Tcr replaced by Kmr from pSKm2 at the BamHI-EcoRV sites | 17 |
| pSKM2 | Apr Kmr; plasmid with the 1.45-kb HindIII-SmaI kanamycin resistance cassette from Tn5 cloned into the HindIII-SmaI sites in pBluescript (SK) | Stellenbosch University laboratory collection (D. E. Rawlings) |