TABLE 1.
Bacterial strains and plasmids
| Bacterial strain or plasmid | Description | Source or reference |
|---|---|---|
| Bacterial strains | ||
| E. coli | ||
| DH5α | supE44 hsdR17 recA1 endA1 gyrA96 thi-1 ΔlacU169 relA1 (φ80lacZΔM15) | BRL |
| JM109λpir | recA1 endA1 gyrA96 thi hsdR17 supE44 relA1 Δ(lac-proAB+) [F′ traD36, proAB lacIqlacZΔM15] λpir | Lab collection |
| V. anguillarum | ||
| 775(pJM1) | Wild type; Pacific Ocean prototype | 19 |
| 531A(pJHC1) | Wild type; Atlantic Ocean prototype | 49 |
| S531A-1 | Plasmidless derivative of 531A | This work |
| 531A(pJHC1δ) | pJHC1 deletion derivative; TAF− | This work |
| 531A(pJHC1::Ω) | pJHC1 derivative containing the Ω transcription-translation terminator at the BglII site in angB; B− G− | This work |
| 531A(pJHC1::K) | pJHC1 derivative containing a Klenow modification at the BglII site in angB; B− G+ | This work |
| 531A(pJHC1::ORFFδ) | pJHC1 derivative containing an in-frame deletion in ORF F | This work |
| Plasmids | ||
| pJM1 | Indigenous plasmid in strain 775 | 17 |
| pJHC1 | Indigenous plasmid in strain 531A | 49 |
| pJHC-S100 | Kmr-encoding cloning vector derived from pBR325 | 37 |
| pBluescript SK | Cloning vector | Stratagene |
| pBR322 | Cloning vector | 8 |
| pTW99 | pBR322 with kanamycin resistance gene from pUC4K cloned into SalI site | This work |
| pTW100 | TAFb region cloned as a BamHI-SalI fragment into pJHC-S100 | This work |
| pTW104 | Derivative of pTW100 containing ORF B as an EcoRV-SpeI fragment | This work |
| pTW105 | pTW104 with the Ω fragment cloned into the BglII site | This work |
| pTW200 | angB and angG cloned by PCR into pTW99 | This work |
| pTW201 | angB cloned by PCR into pTW99 | This work |
| pTW202 | EcoRV-BglII deletion derivative of pTW200 | This work |
| pTW203 | pTW200 containing a Klenow modification at the BglII site in angB | This work |
| pTW200S248L | pTW200 containing an S248L mutation introduced by site-directed mutagenesis | This work |
| pTW203S248L | pTW203 containing an S248L mutation introduced by site-directed mutagenesis | This work |
| pQE60 | T5 phage promoter expression vector | Qiagen |
| pQE60-B+ G+ | angB and angG cloned into pQE60 | This work |
| pQE60-B− G+::K | pQE60-B+ G+ containing a Klenow modification at the BglII site in angB | This work |