TABLE 3.
Transformation efficiency and stability of pLPV plasmids and antibiotic concentrations required for their selection
| Strain (plasmid) | TE (CFU/μg DNA)a | Stability (NAnt/N0)b | Selected by: |
|
|---|---|---|---|---|
| Gm (μg/ml) | Zeo (μg/ml) | |||
| E. coli DH5α(pLPV1Z) | (1.16 ± 0.38) × 105 | 1.07 ± 0.09 | 10 | 25 |
| E. coli DH5α(pLPV2Z) | (1.38 ± 0.31) × 105 | 1.12 ± 0.14 | 10 | 25 |
| E. coli DH5α(pLPV3Z) | (1.80 ± 0.01) × 105 | 1.13 ± 0.11 | 10 | 25 |
| E. coli DH5α(pVRL1) | (1.02 ± 0.21) × 105 | 1.00 ± 0.33 | 10 | |
| A. baumannii ATCC 19606T(pLPV1Z) | (1.56 ± 0.06) × 102 | 0.80 ± 0.23 | 100 | 250 |
| A. baumannii ATCC 19606T(pLPV2Z) | (2.87 ± 0.93) × 102 | 0.83 ± 0.10 | 100 | 250 |
| A. baumannii ATCC 19606T(pLPV3Z) | (4.57 ± 0.29) × 102 | 0.81 ± 0.09 | 100 | 250 |
| A. baumannii ATCC 19606T(pVRL1) | (6.95 ± 0.21) × 102 | 0.92 ± 0.29 | 100 | |
| A. baumannii ATCC17978(pLPV1Z) | (4.37 ± 0.07) × 101 | 0.85 ± 0.14 | 100 | 250 |
| A. baumannii ATCC17978(pLPV2Z) | (5.01 ± 5.01) × 101 | 0.78 ± 0.21 | 100 | 250 |
| A. baumannii ATCC17978(pLPV3Z) | (3.89 ± 0.29) × 101 | 0.75 ± 0.17 | 100 | 250 |
| A. baumannii ATCC17978(pVRL1) | (4.95 ± 0.21) × 101 | 0.92 ± 0.29 | 100 | |
| A. baumannii ACICU(pLPV1Z) | (1.06 ± 0.87) × 102 | 0.74 ± 0.09 | 200 | 250 |
| A. baumannii ACICU(pLPV2Z) | (9.12 ± 0.74) × 103 | 0.73 ± 0.13 | 200 | 250 |
| A. baumannii ACICU(pLPV3Z) | (9.49 ± 1.25) × 103 | 0.71 ± 0.22 | 200 | 250 |
| A. baumannii ACICU(pVRL1) | (1.78 ± 0.21) × 103 | 0.83 ± 0.34 | 200 | |
| A. baumannii AYE(pLPV1Z) | (1.02 ± 1.05) × 103 | 0.92 ± 0.08 | NS | 250 |
| A. baumannii AYE(pLPV2Z) | (1.19 ± 0.98) × 103 | 1.02 ± 0.12 | NS | 250 |
| A. baumannii AYE(pLPV3Z) | (2.84 ± 0.64) × 103 | 0.90 ± 0.11 | NS | 250 |
| A. baumannii AYE(pVRL1Z) | (1.02 ± 0.21) × 103 | 0.95 ± 0.01 | NT | 250 |
| A. pittii UKK_0145(pLPV1Z) | (1.81 ± 0.76) × 104 | 0.67 ± 0.13 | 50 | 250 |
| A. pittii UKK_0145(pLPV2Z) | (1.97 ± 0.04) × 104 | 0.77 ± 0.20 | 50 | 250 |
| A. pittii UKK_0145(pLPV3Z) | (2.84 ± 0.64) × 104 | 0.87 ± 0.17 | 50 | 250 |
| A. pittii UKK_0145(pVRL1) | (1.55 ± 0.05) × 104 | 0.79 ± 0.25 | 50 | |
| A. baylyi BD413(pLPV1Z) | (1.16 ± 0.03) × 104 | 0.64 ± 0.25 | 10 | 25 |
| A. baylyi BD413(pLPV2Z) | (1.38 ± 0.02) × 104 | 0.62 ± 0.27 | 10 | 25 |
| A. baylyi BD413(pLPV3Z) | (1.80 ± 0.01) × 104 | 0.79 ± 0.32 | 10 | 25 |
| A. baylyi BD413(pVRL1) | (9.5 ± 0.75) × 103 | 0.99 ± 0.17 | 10 | 25 |
TE, transformation efficiency. E. coli chemically competent cells were transformed by a heat shock protocol (63); A. baumannii and A. pittii electrocompetent cells were transformed by electroporation (74); A. baylyi naturally competent cells were transformed as previously reported (64). Data are the means ± standard deviations from three independent experiments.
In vitro stability in the absence of antibiotic selection is expressed as the ratio between the number of bacterial colonies grown on LB agar supplemented with the indicated antibiotic concentrations (NAnt) and the number of bacterial colonies grown on LB agar without antibiotic (N0). Data are the means ± standard deviations from three independent experiments. NS, not selectable (Gm MIC > 512 μg/ml); NT, not tested.