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Mutant colonies appeared on selection plates after about 3 days.
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Thus, the mutant colonies appearing under selection may reflect preexisting small-effect mutants that adapted and improved during colony growth.
The overall process looks like time-dependent mutagenesis of resting cells because mutant colonies appear late above a nongrowing lawn.
In spots initiated by single mutants, the purO colonies appeared after 72 hr of incubation and the purR colonies after 96 hr.
Mutant colonies typically appeared 3 to 4 days later than wild-type colonies on 7H11-OADC, and 1 to 2 days later than mc2155/pVV16MSMEG_5084 colonies.
To study mutation under more natural selection conditions, a series of nonlethal systems have been used that prevent growth of the parent cells on selective plates, but still allow mutant colonies to appear over time.
The sptCΔ strains were slightly more impaired in their growth rate, and colonies appeared to be more compact than acdX mutant strains.
We found, however, that when the sea3 Δ mutants that were originally plated on galactose were re-plated on galactose, quantifiable colonies appeared sooner, suggesting that the delay in colony formation in the sea3 Δ mutant was not simply due to a sensitivity to galactose.
No colonies appeared as a result of transfection with polymerase-defective mutants (data not shown).
Wherever new colonies appeared, trouble followed.
Four days after the neomycin-resistant colonies appeared.
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