As the first step in the production of nucleic acid derivatives by microorganisms, adenineless mutants were derived from Brevibacterium ammoniagenes ATCC6872. A culture of Br. ammoniagenes was exposed to ultraviolet rays for 120 second and treated with diethylsulfate in phosphate buffer for 2 hours to reach the designed death rate. The yield of mutants induced was 0.28% by the ultraviolet irradiation and 0.66% by the diethylsulfate treatment. By the diethylsulfate treatment. By the treatment of penicillin G in a hypertonic minimal medium, the yield of mutants was increased from 0.28% to 0.54% and from 0.66% to 1.5%, respectively. Thus, in was demonstrated that diethylsulfate treatment was much more efficient than UV irradiation to induce adenineless mutants of the bacteria, and total strains of 120 adenineless mutants were obtained.