Completed Chromosomes in Thymine-Requiring Bacillus subtilis Spores

Abstract

Origin:terminus genetic marker ratios (both purA: metB and purA:ilvA ) were measured in extracts of spores of Bacillus subtilis strains W23 thy his and 168 thy . For strain W23 thy his , normalized to W23 spore deoxyribonucleic acid, both ratios were equal to unity and were consistent with the presence of only completed chromosomes in the spores. The same ratios in extracts of spores of 168 thy , normalized to strain 168 or the prototroph SB19, were abnormal, i.e., 2.26 ± 0.10 and 0.71 ± 0.06 for purA:metB and purA:ilvA , respectively. These values were unaffected by the extent of extraction of the spore deoxyribonucleic acid, the richness of the medium on which they are formed, and the thymine phenotype. The high ratio for purA:metB is in agreement with the results of earlier workers but, because of the low purA:ilvA ratio, cannot be explained simply by the presence of partially replicated chromosomes in spores of strain 168 thy . Furthermore, purA:leuA in such extracts is 1.01 ± 0.06, consistent with the presence of only completed chromosomes. It is concluded that the abnormal origin:terminus marker ratios are only apparent and result from non-isogenicity between strains 168 thy and 168 in the metB thyB ilvA chromosome region introduced during construction of 168 thy by transformation of strain 168 with W23 thy deoxyribonucleic acid. It is concluded further that the chromosomes of strain 168 thy spores are in a completed form.