Immediately after pulsing, the cells were transferred to 0.5 L prewarmed ammonia medium and grown for 24 h under non-selective conditions at 30°C and 100 rpm shaking.
Thus, the frequency of plasmid loss was less than 1% after seven passages (more than 50 generations) under non-selective conditions in LB medium for the both obtained plasmids RSFmob and RSFmob-I, as well as for RSF1010.
For the both plasmids, the frequency of plasmid loss was less than 1% after four 12-hour passages (more than 50 generations) under non-selective conditions in LB medium.
Yeast strains were grown at 30 °C and 180 rpm in an orbital shaker under non-selective conditions in Yeast Peptone Dextrose (YPD) medium containing 10 g L−1 yeast extract, 20 g L−1 peptone and 20 g L−1 d-glucose.
Under non-selective conditions, yeast strains were grown in YPD medium containing 10 g/L yeast extract, 20 g/L peptone and 20 g/L glucose.
Over 97% of the transformants retained the sulfonylurea resistance phenotype under non-selective conditions.
The Cm-resistant phenotype of the final integrants was stable for more than 50 generations under non-selective conditions.
Strain MSR33 maintained stably the plasmid pTP6 over 70 generations under non-selective conditions.
Therefore, the stability of the plasmid pTP6 in the transconjugant strain MSR33 was studied under non-selective conditions.
All colonies of strain MSR33 grown under non-selective pressure after 70 generations, maintained their mercury resistance.
