TABLE 1.
Strains and plasmids used in this work
| Strain or plasmid | Characteristics and/or application | Antibiotic resistancea | Reference or source |
|---|---|---|---|
| Strains | |||
| 042 | Wild-type Peruvian O44:H18 EAEC isolate | Sm, Tet, Cm | 35, 36 |
| MG1655 | E. coli K-12 strain used as a negative control in HEp-2 adherence assays | E. coli stock center (6) | |
| EC100D pir-116 | Host strain for ori6K plasmids | Epicentre (32) | |
| SB1 | 042Δhra1 isogenic mutant; hra1::aphA-3 | Km, Sm, Tet, Cm | This study |
| SM10 λpir | Donor strain used for conjugal transfer of pSB1 into 042 | Km | 47 |
| thithr leuB tonA lacY supE recA::RP4-2-Tc::Mu-Km | |||
| TOP10 | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74deoRrecA1araD139Δ(ara-leu)7697galUgalKrpsL (Strr) endA1nupG | Invitrogen | |
| OP50 | Nonpathogenic E. coli strain used for C. elegans maintenance | 50 | |
| PA14 | Pseudomonas aeruginosa | 50 | |
| Plasmids | |||
| pBJ1 | hra1 cloned into SspI and SphI sites of pBR322 | Amp | This study |
| pBJ2 | 395-bp central region of hra1 gene deleted from pBJ1 | Amp | This study |
| pBJ4 | pBJ1 hra1::aphA-3; 395-bp central region of hra1 gene in pBJ1 replaced with aphA-3 cassette from pUC18K | Amp, Km | This study |
| pBR322 | Cloning vector | Amp, Tet | New England Biolabs |
| pCVD442 | Suicide vector oriR6K sacB | Amp | 16 |
| pGEMT | TA cloning vector | Amp | Promega |
| pSB1 | aphA-3 cassette and hra1 flanking regions (hra1::aphA-3) in pCVD442 | Amp, Km | This study |
| pTHra1 | hra1 from 042 cloned into pGEMT | Amp | This study |
| pUC18K | pUC18 containing an aphA-3 cassette preceded by translational stop codons in all three reading frames and followed by a consensus ribosomal binding site | 31 |
a
Amp, ampicillin; Km, kanamycin; Sm, streptomycin; Cm, chloramphenicol; Tet, tetracycline.