| | Resistance frequency | Analyses of exconjugants |
|---|
Techniquea | Spontaneousb | Non-spontaneousc | Sampled | Transpositione | Cointegratesf |
|---|
Mating | Not detectable | 1.8 ± 0.53 × 10-3 | 200 | 200 | 0 |
Electroporation | Not detectable | 1.02 ± 0.38 × 10-7 | 100 | 100 | 0 |
- a The pBAM1 plasmid was introduced into recipient cells either by five-hour tri-parental mating or by electroporation, letting the cells to recover after the electro-pulse in LB at 30°C for one hour. Electroporation figures are the average of the frequencies obtained using 100 ng (1.1 ± 0.5 × 10-7) and 500 ng (0.89 ± 0.2 × 10-7) of plasmid DNA.
- b Number of P. putida KT2440 colonies that acquire the marker resistance spontaneously, without mating or electroporation.
- c Total number of cells that acquired the mini-transposon, as measured by growth in kanamycin normalized to the total 3 × 107 donor cells. The 5 h mating frequency was averaged using a total of 16 independent experiments. Electroporation was referred to a final cell concentration of 6 × 1010 electrocompetent cells and the frequency determined with 6 independent experiments.
- d Number of independent colonies that were screened for the presence of the mini-transposon marker (kanamycin) and for the loss of the plasmid backbone marker (ampicillin).
- e Number of kanamycin resistant colonies.
- f Colonies that are both resistance to kanamycin and ampicillin, meaning co-integration of the pBAM1 plasmid into their genome.
