Table 2.
Bacterial strains, plasmids and primers used in this study.
| Strain, plasmid or primer | Relevant characters or sequences (5′–3′)* | Reference or source |
|---|---|---|
| Strain | ||
| E. amylovora | ||
| Ea1189 | Wild‐type, isolated from apple | Burse et al. (2004) |
| Z3205ΔrcsB | rcsB::Km; KmR‐insertional mutant of rcsB of Ea1189, KmR | This study |
| Z3207ΔrcsC | rcsC::Km; KmR‐insertional mutant of rcsC of Ea1189, KmR | This study |
| Z3206ΔrcsD | rcsD::Km; KmR‐insertional mutant of rcsD of Ea1189, KmR | This study |
| Z3205‐06ΔrcsBD | rcsBD::Km; KmR‐insertional mutant of rcsDB of Ea1189, KmR | This study |
| Z0118Δams | KmR‐insertional mutant of ams operon (15.8 kb) of Ea1189, KmR | Zhao and Sundin (2008) |
| Es. Coli | ||
| DH10B | F− mcrA Δ(mrr‐hsdRMS‐mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 endA1 araΔ139 Δ(ara, leu)7697 galU galK λ—rpsL (StrR) nupG | Invitrogen (Carlsbad, CA, USA) |
| Plasmid | ||
| pKD46 | ApR, PBAD gam bet exo pSC101 oriTS | Datsenko and Wanner (2000) |
| pKD13 | KmR, FRT cat FRT PS1 PS2 oriR6 K rgbN | Datsenko and Wanner (2000) |
| pGEM® T‐easy | ApR, PCR cloning vector | Promega |
| pFPV25 | ApR, GFP‐based promoter trap vector containing a promoterless gfpmut3a gene | Valdivia & Falkow (1997) |
| pWSK29 | ApR; cloning vector, low copy number | Wang and Kushner (1991) |
| pWDP1 | 4.54‐kb PCR fragment containing rcsBD gene in pGEM T‐easy vector | This study |
| pWDP2 | 3.88‐kb PCR fragment containing rcsC gene in pGEM T‐easy vector | This study |
| pWDP3 | 4.54‐kb EcoRI‐BamHI fragment containing rcsBD gene in pWSK29 | This study |
| pWDP4 | 721‐bp KpnI‐XbaI DNA fragment containing promoter sequence of amsG gene in pFPV25 | This study |
| pSN1 | 906‐bp EcoRI‐BamHI DNA fragment containing promoter sequence of flhD gene in pFPV25 | This study |
| Primer† | ||
| B3205F | ATGTTGCCATATAAATTTCCGCTAACTTCCGGCAATGTAACACGATTTTTGCGATTGTGTAGGCTGGAGCT | |
| B3205R | TTATTGGTTACCTTGCTGCAGGAGATCTTTGACATAAGTGTCAACTTCACATTCCGGGGATCCGTCGACC | |
| B3206F | GTGAAGTTGACACTTATGTCAAAGATCTCCTGCAGCAAGGTAACCAATAAGCGATTGTGTAGGCTGGAGCT | |
| B3206R | TTATTTATCTACCGGCGTCATGCTTACTGATGACAGGTAGTTAAGCAAAGATTCCGGGGATCCGTCGACC | |
| B3207F | ATGGCCGGGATAACTGGCTTTTTCTGCCTCCCGGCGCTACTGTATCGGCAGCGATTGTGTAGGCTGGAGCT | |
| B3207R | TTACCGCAACCTGTTACTTAGCGCTTCCCGGCTCTTTCTTACCCGCCCGGATTCCGGGGATCCGTCGACC | |
| GFP1 | ATGAGTAAAGGAGAAGAACTTTTCAC | |
| GFP2 | TTATTTGTATAGTTCATCCATGC | |
| AmsG1 | CGGGGTACCGTATCGCTTAAAGGGGAAACG (KpnI) | |
| AmsG2 | CTAGTCTAGACACCTGGAAAGCCATTAATCA (XbaI) | |
| flhD1 | CCGGAATTCCGTTGTTGCCGATGCTAATA (EcoRI) | |
| flhD2 | CGCGGATCCCAATACCCAGGCGAAACATC (BamHI) | |
| RcsC1 | GATCAAGGTAACCGAGCGTAAC | |
| RcsC2 | AGCTTAACCGCAGCATTAAAAC | |
| RcsBD1 | CCGGAATTCCTAGCACAATTCACAAGGTTGG (EcoRI) | |
| RcsBD2 | CGCGGATCCCTCCTAATGAACTGCCGCTACT (BamHI) | |
| Km1 | CAGTCATAGCCGAATAGCCT | |
| Km2 | CGGTGCCCTGAATGAACTGC | |
| Z3205F | GCCAACGCCCGACTGATTACAAG | |
| Z3206R | GCCGGGAAGCGCTAAGTAACAGGT | |
| Z3207F | GTTCGCCGCTAAGCCGTGGTCA | |
| Z3207R | GAGAAAATCAGCGCGGGTGGTTAC | |
*
KmR, ApR, kanamycin and ampicillin resistance, respectively.
†
Italic nucleotides are restriction sites added and the restriction enzymes are indicated at the end of the primers.