TABLE 3.
Strains, plasmids, and primers used in this study
| Strains, plasmids, or primers | Characteristics |
|---|---|
| Strains, and plasmids | |
| Yb2 | Riemerella anatipestifer serotype 2 strain |
| Yb2Δ02955 | AS87_RS02955 gene mutant of R. anatipestifer Yb2 |
| cYb2Δ02955 | Mutant Yb2Δ02955 carrying plasmid pCP29-AS87_RS02955 |
| pCP29 | ColE1 ori (pCP1 ori), Apr (Emr); E. coli-F. johnsoniae shuttle plasmid |
| pCP29-AS87_RS02955 | pCP29 containing ompA promoter and AS87_RS02955 ORF,a cfxAb (Apr) |
| pCold I | ColE1 ori, Apr, N-terminal His-tag |
| Primers | |
| AS87_RS02955-F | 5′-GGTACAGGTGGTAATGATGG-3′ |
| AS87_RS02955-R | 5′-GAAACTTTCTCACCGTTTAC-3′ |
| AS87_RS02955 P1-F | 5′-CATAATGCACAACGGGTAATAACTTTA-3′ |
| AS87_RS02955 P1-R | 5′-GAACGGGCAATTTCTTTTTTGTCATGAGAGAGTAATTTTTTCATTTTTAA-3′ |
| Erm-P-F | 5′-TTAAAAATGAAAAAATTACTCTCTCATGACAAAAAAGAAATTGCCCGTTC-3′ |
| Erm-P-R | 5′-AAAAATAAATTTGACTTATTTCTTGCTACGAAGGATGAAATTTTTCAGGG-3′ |
| AS87_RS02955 P2-F | 5′-CCCTGAAAAATTTCATCCTTCGTAGCAAGAAATAAGTCAAATTTATTTTT-3′ |
| AS87_RS02955 P2-R | 5′-TTGTTTTTGTACTTGCTCAAAATGATT-3′ |
| Erm-F | 5′-ATGACAAAAAAGAAATTGCCCGTT-3′ |
| Erm-R | 5′-CTACGAAGGATGAAATTTTTCAGGG-3′ |
| 16S rRNA-F | 5′-GAGCGGTAGAGTATCTTCGGATACT-3′ |
| 16S rRNA-R | 5′-TATTACCGCGGCTGCTGGCA-3′ |
| AS87_RS02955-orf-F | 5′-CGACTCGAGATGAAAAAATTACTCTCTC-3′ (XhoI site underlined) |
| AS87_RS02955-orf-R | 5′-CATGCATGCTTATTTCTTGATAATTTTTTGAG-3′ (SphI site underlined) |
| AS87_RS02955-pCold-F | 5′-TCGGTACCCTCGAGGGATCCTGCATCAATGCTCAAGTTTTC-3′ |
| AS87_RS02955-pCold-R | 5′-TCTAGACTGCAGGTCGACTTTCTTGATAATTTTTTGAG-3′ |
| Chβ-actin-F | 5′-GAGAAATTGTGCGTGACATCA-3′ |
| Chβ-actin-R | 5′-CCTGAACCTCTCATTGCCA-3′ |
| ChTNF-α-F | 5′-CGCTCAGAACGACGTCAA-3′ |
| ChTNF-α-R | 5′-GTCGTCCACACCAACGAG-3′ |
| ChIL-1β-F | 5′-GGGCATCAAGGGCTACA-3′ |
| ChIL-1β-R | 5′-TCGGGTTGGTTGGTGATG-3′ |
| ChIL-6-F | 5′-GTTCGCCTTTCAGACCTAC-3′ |
| ChIL-6-R | 5′-ACCACTTCATCGGGATTTA-3′ |
| ChIL-8-F | 5′-TTGGAAGCCACTTCAGTCAGAC-3′ |
| ChIL-8-R | 5′-GGAGCAGGAGGAATTACCAGTT-3′ |
| AS87_RS02955-F22-F | 5′-TCGGTACCCTCGAGGGATCCTGCATCAATGCTCAAGTTGCCCAAG-3′ |
| AS87_RS02955-F26-F | 5′-TCGGTACCCTCGAGGGATCCTGCATCAATGCTCAAGTTTTCCAAGAAAACGCTAATGACATTAATGG-3′ |
| AS87_RS02955-F39-F | 5′-GCTAGTGGTAATGTAGCCTCTGCACCTCTCAATTCTTA-3′ |
| AS87_RS02955-F39-R | 5′-GAGGCTACATTACCACTAGCAACACCATCATTACCACCT-3′ |
| AS87_RS02955-W54-F | 5′-GCGACTCTAAACAGAACTTACGAAGCTAATGCGTGTATTA-3′ |
| AS87_RS02955-W54-R | 5′-CGTAAGTTCTGTTTAGAGTCGCGTCTCCATAAGAATTGAG-3′ |
| AS87_RS02955-L90-F | 5′-GCTTCCTTTAAAGCAGCCGCTTGGGATACTAAAACAGAA-3′ |
| AS87_RS02955-L90-R | 5′-GCGGCTGCTTTAAAGGAAGCTTTTCCGTTTCCTGTTAAT-3′ |
| AS87_RS02955-F92-F | 5′-GCTAAAGCAGCCGCTTGGGATACTAAAACAGAACAAACCT-3′ |
| AS87_RS02955-F92-R | 5′-CAAGCGGCTGCTTTAGCGGAAAGTTTTCCGTTTCCTGTTA-3′ |
| AS87_RS02955-L106-F | 5′-GCTAAAATTTCAGCAACTGGAGGAACTCTAAGTGTTTCAGA-3′ |
| AS87_RS02955-L106-R | 5′-GTTGCTGAAATTTTAGCAGAGGTTTGTTCTGTTTTAGTATCCC-3′ |
| AS87_RS02955-L122-F | 5′-GCAGACAAAGGAGCTTTTAAGATCTATACAGTAGATATCACTAACG-3′ |
| AS87_RS02955-L122-R | 5′-AAAGCTCCTTTGTCTGCTGTAACTTCTGAAACACTTAGAGTTC-3′ |
| AS87_RS02955-Y130-F | 5′-GCTACAGTAGATATCACTAACGCAACGGGAGATTTAAAG-3′ |
| AS87_RS02955-Y130-R | 5′-GCGTTAGTGATATCTACTGTAGCGATCTTAAAAGCTCC-3′ |
| AS87_RS02955-I134-F | 5′-GCCACTAACGCAACGGGAGATTTAAAGATTACCTTTGAAGG-3′ |
| AS87_RS02955-I134-R | 5′-CTCCCGTTGCGTTAGTGGCATCTACTGTATAGATCTTA-3′ |
| AS87_RS02955-F145-F | 5′-GCTGAAGGTTTTCAGGCTGCTAGATCAAGATTTTTTTTGGACG-3′ |
| AS87_RS02955-F145-R | 5'-GCAGCCTGAAAACCTTCAGCGGTAATCTTTAAATCTCC-3′ |
| AS87_RS02955-F155-F | 5′-GCTTTTTTGGACGATATAGTAGTTACAGAAACCACTATGTCTG-3′ |
| AS87_RS02955-F155-R | 5′-CTATATCGTCCAAAAAAGCTCTTGATCTAGCAGCCTGAAAACC-3′ |
a
ORF, open reading frame.
b
cfxA, cefoxitin resistance gene.