SYNERGISTIC KILLING EFFECT OF GLYCINE AND PENICILLIN, AND RAPID METHODS FOR MUTANT ISOLATION IN PSEUDOMONAS

Abstract
The addition of 3% glycine and 10 units/ml of penicllin-G to a culture of Pseudomonas resulted in rapid death. More than 95% of viable cells were killed within 30 min of incubation. Cell wall synthesis, as determined byamino-acid incorporation, was inhibited by glycine plus penicillin. Penicillin in high concentrations induced some kind of unbalanced growth, while EDTA which had little or no effect on intact cells or cells treated with penicillin in a concentration of 1x10-4[image] lysed cells treated with glycine and penicillin. The combined action of these chemicals was utilized for the rapid screening of temperature-sensitive mutants from UV-irradiated cells. N-Nitroso-N-methylurethane was an extremely efficient mutagen for the isolation of temperature-sensitive mutants. The frequency of mutation was so high that mutants can be isolated without any screening procedure.

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