TABLE 1.
Strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Characteristicsa | Source or reference |
|---|---|---|
| Strains and plasmids | ||
| Yb2 | Riemerella anatipestifer serotype 2 strain | 4 |
| Escherichia coli S17-1 | λpir hsdR pro thi; chromosomally integrated RP4-2, Tc::Mu Km::Tn7 | 18 |
| RA625 | Tn4351 insertion mutant of R. anatipestifer Yb2, pncA::Tn | 12 |
| cYb2ΔpncA | Mutant RA625 carrying plasmid pCP29-pncA | This study |
| pCP29 | ColE1 ori (pCP1 ori), Apr (Emr); E. coli-F. johnsoniae shuttle plasmid | 18 |
| pCP29-pncA | pCP29 containing ompA promoter and pncA ORF, cfxA (Apr) | This study |
| Primers | ||
| ompA promoter P1 | 5′-CAGGTACCATAGCTAAAATTTTGGCAGTAAC-3′ (KpnI site underlined) | 18 |
| ompA promoter P2 | 5′-CGACTCGAGCATTCCAATTCTCTTATTATC-3′ (XhoI site underlined) | 18 |
| pncAorf-F | 5′-CCGCTCGAGATGAAAAAAGCACTTATCGTGG-3′ (XhoI site underlined) | This study |
| pncAorf-R | 5′-CATGCATGCTTAACCTAAAATGGTAACACCA-3′ (SphI site underlined) | This study |
| PncA-F | 5′-CGGGATCCATGAAAAAAGCACTTATCGTGGT-3′ (BamHI site underlined) | This study |
| PncA-R | 5′-CCCAAGCTTACCTAAAATGGTAACACCATTT-3′ (HindIII site underlined) | This study |
a
ORF, open reading frame; cfxA, cefoxitin resistance gene.