Table 1.
Bacterial strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Description or sequence (5′–3′)a | Source or reference |
|---|---|---|
| Strains | ||
| V. salmonicida strains | ||
| LFI1238 | Wild type (wt); isolated from Atlantic cod | 26 |
| ΔlitR | LFI1238 containing an in-frame deletion in litR | This study |
| ΔlitRc | ΔlitR strain complemented with the LFI1238 litR gene; Cmr | This study |
| E. coli strains | ||
| S17-1 | Donor strain for conjugation; λ pir | 60 |
| β-2855 | Donor strain for conjugation; diaminopimelic acid auxotroph; λ pir | 34 |
| Plasmids | ||
| pDM4 | Cmr; suicide vector with an R6K origin (λ pir requiring) and sacBR | 45 |
| pNQ705 | Cmr; suicide vector with an R6K origin (λ pir requiring) | 45 |
| pDM4ΔlitR | pDM4 containing a fragment of litR harboring an internal deletion | This study |
| pNQ705litR+ | pNQ705 containing wt litR and flanking sequences | This study |
| Primers | ||
| Primers for construction of ΔlitR and ΔlitRc strains | ||
| LitR-A fwd | CCGCTCGAGTAGTTCCATAATCTTTTCTATG | This study |
| LitR-B rev | CTTACTCTACTTATATTTATTATATCCTTGCCAAC | This study |
| LitR-C fwd | TATAAGTAGAGTAAGCGTGC | This study |
| LitR-D rev | GGACTAGTGTTATTTTCGGGTTCAAC | This study |
| Primers for verification of plasmids, transconjugants, and mutants | ||
| LitR-G fwd | ACCAACGGCAGGACTTAGAC | This study |
| LitR-H rev | TTGATAACAATCGAGCAGAGC | This study |
| Pnq-fwd | TAACGGCAAAAGCACCGCCGGACATCA | Debra Miltonb |
| Pnq-rev | TGTACACCTTAACACTCGCCTATTGTT | Debra Milton |
| LitR-fwd | GTGTGGTTTGAATGGAGCAC | This study |
| LitR-rev | CTCAATCCCTTCAGCAAACA | This study |
| qRT-PCR primers | ||
| LitR-fwd | GTGTGGTTTGAATGGAGCAC | This study |
| LitR-rev | CTCAATCCCTTCAGCAAACA | This study |
| 16S-fwd | CTTGACGTTAGCGACAGAAGAA | This study |
| 16S-rev | CGCTTTACGCCCAGTAATTC | This study |
| AccD-fwd | TTGCTGGTCGTCGTGTTATT | This study |
| AccD-rev | TTTAGCCATCAAACCACCAA | This study |
| FtsZ-fwd | CGGATGTTCGTACGGTAATG | This study |
| FtsZ-rev | CAAGCAATGGGCTTGAGATA | This study |
| RpoD-fwd | AAGCCGAAGAAATTCGCTTA | This study |
| RpoD-rev | GCAAGATCTGATTCGCTCAA | This study |
a
Cmr, chloramphenicol resistance.
b
Umeå University, Sweden.