TABLE 2.
Plasmids used in this study
| Plasmid | Description | Source or Ref. |
|---|---|---|
| pGP704 | Mobilizable suicide vector, R6K2 replicon, requires π proteins in trans from λpir positive strain | (47) |
| pUC-A-1 | pUC13 with 5 kb EcoRI/HindIII fragment of pYVO8 from WA-314, carrying yadA gene | (23) |
| pUC-SS | pUC-A-1 derivative, carrying the yadA signal sequence (bp 1–87) | This study |
| pGP-SS | pGP704 with EcoRI/SphI fragment of pUC-SS | This study |
| pGPS-SS | 1,8 kb spectinomycin (spcR) resistance cassette in EcoRI site of pGP-SS | This study |
| pGPS-nadA | pGPS-SS with nadA MC58 bp 75–1095 | This study |
| pYVO8-A-0 | pYVO8, yadA negative, kmR resistance cassette inserted in PstI site of yadA by allelic exchange | (23) |
| pYVO8-SS | pYVO8-A-0 with integrated pGPS-SS | This study |
| pYVO8-A-1 | pYVO8-A-0 with integrated pGPS-A-1, wild type yadA | (23) |
| pYVO8-nadA | pYVO8-A-0 with integrated pGPS-nadA | This study |
| pACYC184 | cloning vector pACYC184 | (49) |
| p | pACYC184 with virF (plus 151 bp upstream and 86 bp downstream of virF) | This study |
| pnadA | p with nadA MC58 (bp 75–1095), yadA promoter (144 bp upstream of yadA), yadA encoding LP (bp 1–90) and yadA terminator (119 bp downstream of yadA) | This study |
| pyadA | p with yadA (bp 1–1266), yadA promoter (144 bp upstream of yadA) and yadA terminator (119 bp downstream of yadA) | This study |
| pET21b+ | Expression with C-terminal His-tag and T7-promotor | Novagen (Darmstadt, Germany) |
| pET21b:nadA | pET21b+ with nadA bp 75–636 | This study |