TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Characteristic(s)a | Reference or source |
|---|---|---|
| P. pseudoalcaligenes strains | ||
| CECT5344 | Wild type; uses cyanide as N source | 24 |
| CECT5344N | Spontaneous mutant resistant to nalidixic acid | This work |
| CECT5344N lacking the cyn gene | Gm-directed mutant in the cynS gene | This work |
| E. coli strains | ||
| DH5α | Lac[minus]; host for most of the plasmids | 32 |
| S17-1 | Tra+; host for the mobilizable mob plasmids | 35 |
| Plasmids | ||
| pGEM-T | Vector (Apr) used for cloning fragments amplified by PCR | Promega |
| pBluescript SK(+) | Multipurpose cloning vector (Apr) | Stratagene |
| pK18mobδE | Suicide vector in Pseudomonas spp. (Kmr) | 33 |
| pMS255 | Vector containing a Gm resistance cassette | 4 |
| pVIC1 | pK18mobδE derivative containing the 1.3-kb fragment of the cynS gene | This work |
| pVIC2 | pVC1 derivative with the cynS gene interrupted by the Gm cassette | This work |
| pMH1 | pBluescript SK(+) harboring the SalI fragment (1.5 kb) that contains the entire cynS gene | This work |
a
Apr, ampicillin resistant; Kmr, kanamycin resistant.