TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Genotype and/or relevant characteristic | Source or reference |
|---|---|---|
| Strains | ||
| A. salmonicida | ||
| AS1102 | Type strain, A-layer negative | NCIMB 1102 |
| 644Rb | Virulent isolate, Nalr, A-layer positive | 35 |
| MT1326 | Virulent isolate, A-layer positive | 3 |
| CB3 | Undefined Tn5 mutant, decreased synthesis of extracellular proteins including GCAT and serine protease | 6 |
| 644RbΔG | ΔsatA::kan, serine protease, A-layer positive | This study |
| 644RbΔSP | ΔaspA::kan, pro-GCAT, A-layer positive | This study |
| MT1326ΔG | ΔsatA::kan, serine protease, A-layer positive | This study |
| MT1326ΔSP | ΔaspA::kan, pro-GCAT, A layer positive | This study |
| E. coli | ||
| HB101 | Δ(gpt-proA)62 leuB6 supE44 ara-14 galK2 lacY1 Δ(mcrC-mrr) rpsL20 xyl-5 mtl-1 recA13 | 1 |
| DH5α | recA1gyrA relA1 Δ(lacIZYA-argF)U169 deoR [φ80dlac lacZΔM15] | 1 |
| S17-1 | thi pro hsdR hsdM+ recA [RP4 2-Tc::Mu-Km::Tn7 (Tpr Smr) Tra+] | 32 |
| Plasmids | ||
| pUC18/19 | Cloning and sequencing | 1 |
| pBSK | Bluescript SKII, cloning and sequencing | Stratagene |
| M13mp18/19 | Sequencing and mutagenesis | 1 |
| pSUP202 | Mobilizable suicide vector (Ampr Tetr Camr ColE1 ori Mob+) | 32 |
| pSM20 | Levansucrase confers sucrose sensitivity (sacRB+ Ampr) | 31 |
| pRAT1 | Kanamycin resistance determinant, lacking transcriptional terminator, on 1.436-kb HaeII fragment from Tn903 | D. Evans, University of Reading |
| pMMB66HE | Broad-host-range expression vector (Ampr) | 17 |
| pSUP202sac | Mobilizable suicide vector, confers Sucs (Mob+sacRB+ Tetr) | This study |
| pUC18G | satA on 1.2-kb PstI fragment, cloned in PstI site of pUC18 in reverse orientation to plac | This study |
| pUC19S | satA on 5.5-kb SacI fragment in SacI site of pUC19 | This study |
| pUC19SΔG | pUC19S with Kan cassette replacing an internal fragment of satA | This study |
| pSUP202sacΔG | SacI fragment from pUC19SΔsatA::kan inserted in pSUP202sac | This study |
| pBSKSP | aspA cloned in pBSK | This study |
| pBSKΔSP | pBSKSP with Kan cassette replacing small internal fragment of ΔaspA | This study |
| pSUP202sacΔSP | ΔaspA::kan on PstI fragment from pBSKSPΔaspA::kan in PstI site of pSUP202sac | This study |
| pMMBG | satA subcloned on 1.2-kb PstI fragment in PstI site of pMMB66HE, under tac control | This study |
| M13mp18G+/− | satA on 1.2-kb PstI fragment in forward or reverse orientation in PstI site of M13mp18 | This study |
| pMMBG228-Ala | pMMBG encoding mutation of Gly-228 to Ala-228 | This study |