TABLE 1.
Reference strains and plasmids used in this study
| Strain or plasmid | Relevant genotype or phenotype | Source or reference |
|---|---|---|
| Strains | ||
| E. coli JM109 | endA1 gyrA96 hsdR17 Δ(lac proA) relA recA1 supE44 thi F (lacIqZΔM15 proAB+traD36) | Gibco BRL-Life Technologies |
| E. coli DH10B | araD139 Δ(ara leu)7697 deoR endA1 galK1 galU nupG recA1-rpsL F′ mrcA Δ(mrr-hsdRMS-mrcBC)Φ80dlacZΔM15 ΔlacX74 | Stratagene |
| Plasmids | ||
| pPCRScript Amp SK(+) | Ampicillin resistant | Stratagene |
| pACYC184 | Chloramphenicol resistant, low copy number | 11 |
| pBK-CMV | Kanamycin resistant, high copy number | Stratagene |
| pILT-3 | Natural ca. 50-kb plasmid of K. pneumoniae ILT-3 producing CTX-M-19 | 39 |
| β-lactamase | ||
| pA-1 | Recombinant pPCR-Script Amp with a 6.5-kb PCR fragment from whole-cell DNA of K. pneumoniae ILT-3 containing ISEcp1B and a truncated blaCTX-M-19 gene | This study |
| pB-1 | Recombinant pPCR-Script Cam with a 1.1-kb PCR fragment from whole-cell DNA of K. pneumoniae ILT-3 containing ISEcp1B promoter sequences and the entire blaCTX-M-19 gene | This study |
| pB-2 | Recombinant pPCR-Script Cam with a 1-kb PCR fragment from whole-cell DNA of K. pneumoniae ILT-3 lacking ISEcp1B-promoting sequences and containing the entire blaCTX-M-19 gene | This study |