Table 1.
Strains and plasmids used in the current study
| Strains or Plasmid | Relevant characteristic(s) | Reference or Source |
|---|---|---|
| Strains | ||
| E. coli RS218 | E. coli RS218 (O18:K1:H7), the cerebrospinal fluid isolate from a neonate with meningitis | (Khan et al., 2002) |
| E. coli EC100D | F− mcrA Δ(mrr-hsdRMS-mcrBC) Φ80dlacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ− rpsL nupG pir+(DHFR). | Epicentre Biotechnologies |
| E. coli DH5α | F′ Phi80dlacZ DeltaM15 Delta(lacZYA-argF)U169 deoR recA1 endA1 hsdR17(rK-mK+)phoA supE44 lambda- thi-1 | Lab stock |
| NBC | β-lactamase reporter gene was translationally fused to the C- terminal of cnf1 gene in the chromosome of strain RS218 | (Yu and Kim, 2010) |
| YK3 | E. coli RS218 (O18:K1:H7) gidA deletion mutant | This study |
| YK4 | E. coli RS218 (O18:K1:H7) gidA deletion mutant compelemented with gidA under the control of its native promoter. Complementation was achieved by Tn7 site- specific insertion into a second benign site in the chromosome. | This study |
| YK05 | E. coli RS218 (O18:K1:H7) gidA deletion mutant, compelemented with N-terminal truncated GidA under the control of arabinose promoter. Complementation was achieved by Tn7 site-specific insertion into a second benign site in the chromosome. | This study |
| YK06 | E. coli RS218 (O18:K1:H7) with additional copy of N- terminal truncated GidA under the control of arabinose promoter. Complementation was achieved by Tn7 site- specific insertion into a second benign site in the chromosome. | This study |
| Plasmids | ||
| pCXN | CNF1 coding region was cloned into the KpnI site of pCX340, tetracycline resistance. | (Yu and Kim, 2010) |
| pKD3 | Containing chromphenicol resistance gene, R6kγ replication origin | (Datsenko and Wanner, 2000) |
| pKD47 | a derivative of pKD46 (Datsenko & Wanner, 2000), with the only modification that blaM in pKD46 was replaced by spectinomycin resistance gene. | (Yu and Kim, 2010) |
| pGRG36 | Tn7 insertion vector, ampicillin resistance, temperature sensitive | (Mckenzie and Craig, 2006) |
| pGAP | Multiple cloning site of pGRG36 was ligated into PvuII site of pBC-KS, resulting in pGRGM. Then AraC and arabinose promoter (pBAD) was cloned into AvrII and XhoI site of pGRGM. | (Yu and Kim, 2010) |
| pCX340 | PBR322 derivative, cloning vector used to fuse CNF1 to the mature form of TEM1 β-lactamase, tetracycline resistance. | (Charpentier and Oswald, 2004) |
| pSR | Tn5 vector, spectinomycin resistance, R6kγ replication origin | This study |
| pGAP- gidAΔN | The coding region of N-terminal truncated GidA was cloned into NdeI and NotI site of pGAP | This study |
| pG-gidAΔN | DNA fragment containing truncated gidA gene that obtained from pGAP-gidAΔN by digestion with AvrII and PacI, and ligated into same sites of pGRG36. | This study |
| pG-gidA | DNA fragment containing gidA gene that obtained by PCR amplification from genomic DNA, and ligated into same sites of pGRG36. | This study |