TRANSFORMATION OF BACILLUS LICHENIFORMIS

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Abstract
When a series of 28 auxotrophic mutants of Bacillus licheniformis were screened for transformation, only three of them, M28 (glycine), M30 (uncharacterized), and M33 (purine), produced a detectable number of transformants. The screening method consisted of spreading auxotrophic cells and deoxyribonucleic acid (DNA) from the prototrophic strain 9945A on minimal agar plates and observing the plates for development of prototrophic colonies. M28 transformed at a higher frequency than did the two other mutants, and it was studied in greater detail. Although up to 20% of the recipient cells spread on the plates in the presence of DNA gave rise to prototrophic colonies over a period of 72 hr, only about 10-3 % of the cells produced transformants when they were incubated with DNA in liquid suspension for 1 hr. The most competent cultures of many tested were those grown on a shaker for 22 hr in a medium composed of nutrient broth, salts, and glycerol. When mutations resulting in requirements for histidine, leucine, serine, and tryptophan were introduced singly into the glycine mutant, transformants for the leucine, serine, and histidine markers could be obtained at will, but transformants for the tryptophan marker were not detected even though all four of the double mutants could be transformed to glycine independence.