Fujita Y, Komano T, Tanooka H
J Bacteriol. 1973 Feb;113(2):558-64. doi: 10.1128/jb.113.2.558-564.1973.
The incorporation of (3)H-labeled thymidine triphosphate ((3)H-dTTP) into deoxyribonucleic acid (DNA) of germinated and then Brij 58-treated Bacillus subtilis spores was measured to study DNA replication activity of cells. The dTTP incorporation rate was very low in dormant spores, gradually increased as germination proceeded, and reached a level of the vegetative cell activity approximately 4 hr after the start of germination. This is in contrast to the DNA polymerase activity in the cell extract which remained at the same level throughout the germination period. The increase of the dTTP incorporation activity was inhibited by chloramphenicol or phenethyl alcohol. When these inhibitors were added after germination had proceeded, the elevated dTTP incorporation activity gradually decreased. Permeability to dTTP of spores germinated in the presence of chloramphenicol and then treated with Brij 58 was confirmed by (i) (3)H-dTTP incorporation into the treated spores following either electron or ultraviolet irradiation and (ii) release of radioactivity from the treated spores containing radioactively labeled DNA after deoxyribonuclease I treatment.
通过测量（³H）标记的三磷酸胸腺嘧啶核苷（（³H）-dTTP）掺入已萌发然后经Brij 58处理的枯草芽孢杆菌孢子的脱氧核糖核酸（DNA）中的情况，来研究细胞的DNA复制活性。在休眠孢子中，dTTP掺入率非常低，随着萌发的进行逐渐增加，并且在萌发开始后约4小时达到营养细胞活性水平。这与细胞提取物中的DNA聚合酶活性形成对比，后者在整个萌发期保持在相同水平。dTTP掺入活性的增加受到氯霉素或苯乙醇的抑制。当在萌发进行后添加这些抑制剂时，升高的dTTP掺入活性逐渐降低。通过以下方法证实了在氯霉素存在下萌发然后用Brij 58处理的孢子对dTTP的通透性：（i）在电子或紫外线照射后，（³H）-dTTP掺入经处理的孢子中；（ii）在脱氧核糖核酸酶I处理后，含有放射性标记DNA的经处理的孢子释放出放射性。
