TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant genotype or phenotypea | Source or reference |
|---|---|---|
| Strains | ||
| E. coli DH10B | F′ mcrA Δ(mrr-hsdRMS mcrBC) φ80dlacZΔM15 ΔlacX74 deoR recA1 araΔ139 Δ(ara leu)7697 galU galK λ− rpsL endA1 nupG Strr | Life Technologies |
| E. coli MG-1 | Extended-spectrum cephalosporin resistance, Rifr Kanr Tobr Genr Amkr Sptr Strr Sulr Tetr Cmr | 51 |
| Plasmidsb | ||
| pPCR-script Amp (SK+) | Ampr | Stratagene |
| pPCR-script Cm (SK+) | Cmr | Stratagene |
| PBK-CMV | Neor Kanr | Stratagene |
| pNLT1 | Natural plasmid of E. coli MG-1 containing blaVEB1 ESBL, Rifr Kanr Tobr Genr Amkr Sptr Strr Sulr Cmr | 51 |
| pRLT2 | pBKCMV recombinant plasmid containing an 11-kb Sau3A fragment with blaVEB-1 ESBL, Kanr Tmr Amkr Netr Cmr Sulr | This work |
| pRLT3 | pBKCMv recombinant plasmid containing a 13-kb Sau3A fragment with blaVEB-1 ESBL, Kanr Tmr Amkr Netr Cmr | This work |
| pRLT4a and b | pPCRscript recombinant plasmid containing a 1.3-kb PCR fragment with qacI; a, EtBrr CTABr; b, Etbrr CTABr | This work |
| pRLT5a and b | pPCRscript recombinant plasmid containing a 1.8-kb PCR fragment with aacA1/orfG; a, Kanr Tmr Amkr Netr; b, no resistance observed | This work |
| pRLT6a and b | pPCRscript recombinant plasmid containing a 2.7-kb PCR fragment with arr-2; a, Rifr; b, no resistance observed | This work |
| pRLT7a and b | pPCRscript recombinant plasmid containing a 3.8-kb PCR fragment with cmlA5; a, Cmr; b, Cmr | This work |
| pRLT8a and b | pPCRscript recombinant plasmid containing a 3.2-kb PCR fragment with oxa10/aadA1; a, Ampr Strr Sptr; b, Ampr Strr Sptr | This work |
| pRLT9a and b | pPCRscript recombinant plasmid containing a 1.9-kb PCR fragment with the oxa10/aadA1 fusion cassette; a, Ampr Strr Sptr; b, Ampr Strr Sptr | This work |
| pRLT10a and b | pPCRscript recombinant plasmid containing a 0.9-kb PCR fragment with the oxa10 cassette; a, Ampr; b, no resistance observed | This work |
a
Abbreviations: Kan, kanamycin; Tm, tobramycin; Net, netilmycin; Str, streptomycin; Spt, spectinomycin; Amk, amikacin; Cm, chloramphenicol; Tet, tetracycline; Sul, sulfonamide; Rif, rifampin; ESBL, extended-spectrum β-lactamase.
b
For plasmids with “a” and “b” designations, “a” indicates recombinant plasmids where the inserted genes are in the same orientation as the Plac promoter of pPCRscript and “b” indicates recombinant plasmids where the inserted genes are in the orientation opposite that of the Plac promoter of pPCRscript.