Table 1.
Strain, Plasmid and Primer
| Strain | Genotype a, b | Source or Reference |
|---|---|---|
| Dickeya dadantii | ||
| EC 3937 | Wild-type | Laboratory collection |
| A 4229 | ompF::uidA-Kan | Condemine and Ghazi, 2007 |
| NFB 7012 | mTn5degP::uidA-Kan | This study |
| NFB 7039 | mTn5spy::uidA-Kan | This study |
| NFB 7515 | cpxA::gm | This study |
| NFB 7532 | cpxR::gm | This study |
| NFB 7539 | cpxA::gm ompF::uidA-Kan | This study |
| NFB 7543 | cpxR::gm ompF::uidA-Kan | This study |
| NFB 7548 | cpxR::gm mini-Tn5degP::uidA-Kan | This study |
| NFB 7549 | cpxA::gm mini-Tn5degP::uidA-Kan | This study |
| NFB 7550 | cpxR::gm mini-Tn5spy::uidA-Kan | This study |
| NFB 7551 | cpxA::gm mini-Tn5spy::uidA-Kan | This study |
| Escherichia coli | ||
| BL21(DE3) | ompT, hsdSB, gal, dcm | Invitrogen |
| Top 10 F’ | F′(lacIq, Tn10) mcrA, Δ(mrr-hsdRMS-mcrBC), Φ80lacZΔM15, ΔlacX74, recA1, araD139, Δ(ara- leu)7697, galU, galK, rpsLendA1 nupG | Invitrogen |
| S17-λpir | recA1, thi, pro, hsdR-M+, RP4:2-Tc::Mu-Kan::Tn7, λpir | De Lorenzo et al., 1994 |
| Plasmids | ||
| pUC18Not-uidA | ‘uidA, Amp | Bontemps-Gallo et al., 2014 |
| pJET 1.2 | Amp | Fermentas |
| pNFW460 | pJET2.1 cpxR | This study |
| pNFW466 | pJET2.1 cpxR::gm | This study |
| pNFW512 | pJET2.1 cpxA | This study |
| pNFW513 | pJET2.1 cpxA::gm | This study |
| pUTmini-Tn5-Kan | mini-Tn5Spe, oriR6K, Kan, Amp | De Lorenzo et al., 1990 |
| Primer sequencesc | ||
| cpxAFor | CGCGAGCTGACATCCCTATT | This study |
| cpxARev | TCGAAAAAGCTCTCCAGCGT | This study |
| cpxRFor | CTATCATCCAGCCCCTGAC | This study |
| cpxRRev | CGGATGCTGTTTCAGGTTAC | This study |
| K7gentabsrFor | TCTCTGTACACCCCCATCCCCCTGTTGAC | This study |
| K7gentabsrRev | TCTCTGTACACGCAAGCTAGCTTGGCTGC | This study |
| K7gentahpaFor | GTGTGTTAACCCCCATCCCCCTGTTGAC | This study |
| K7gentahpaRev | GTGTGTTAACCGCAAGCTAGCTTGGCTGC | This study |
| spyForKpnI | AAGGTACCTGATGGCTCCTGCCGCCGGCGA | This study |
| spyRevXbaI | GTTCTAGAAGCCGACGCTACCCAGCGCCAG | This study |
| degPForKpnI | TTGGTACCACAAACTCTCCAGCAAGCATTG | This study |
| degPRevXbaI | TGCTCTAGACTCAACGCCAACGCACTCAGC | This study |
| cpxRhisFor | CACCATGAACAAAATCCTGTTGGTTGATGACG | This study |
| cpxRhisRev | CGGTAACCCGAATCATGCGGTGG | This study |
a
The degP-uidA, cpxP-uidA and spy-uidA gene fusions are carried by a mini-Tn5 Kan
b
Amp, ampicillin resistance, Kan, kanamycin resistance, Gm, gentamicin resistance
c
Restriction sites are underlined