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Haemin dependent mutants were isolated from Bacillus subtilis after (a) ultraviolet irradiation and selection by streptomycin, (b) exposure of the organisms to copper ions and (c) treatment with N-methyl-N′-nitro-N-nitro-soguanidine. The isolates were grown in a synthetic medium containing haemin when substances able to neutralize inhibitory factors and to stabilize the medium were included. Two of the mutants were capable of growth in media supplemented only with δ-aminolaevulinic acid, the first intermediate of porphyrin synthesis. Transformation of the mutants to haemin independence was accomplished using deoxyribonucleic acid.
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