Sonnenfeld E M, Koch A L, Doyle R J
J Bacteriol. 1985 Sep;163(3):895-9. doi: 10.1128/jb.163.3.895-899.1985.
The origin of replication of Bacillus subtilis 168 trp thy dna-1 (temperature-sensitive initiation mutant) was labeled with [3H]thymidine. Analysis of labeled cells by autoradiography revealed that most of the radioactivity was associated with cell pole areas. To label the terminus, cells that had initiated were treated with chloramphenicol to inhibit cell growth and division but to allow continued DNA synthesis. These cells were then labeled with [3H]thymidine at a time when chromosome replication was nearly complete. The distribution of radioactivity was similar to that observed in origin-labeled cells. In contrast, exponentially growing cells that were labeled for a brief time at the permissive temperature showed a random distribution of radioactivity. These data indicate that the origin and terminus of replication are located at cell poles.
用[3H]胸苷标记枯草芽孢杆菌168 trp thy dna - 1(温度敏感起始突变体)的复制起点。通过放射自显影对标记细胞进行分析,结果显示大部分放射性与细胞极区相关。为了标记复制终点,对已起始复制的细胞用氯霉素处理,以抑制细胞生长和分裂,但允许DNA继续合成。然后在染色体复制几乎完成时,用[3H]胸苷对这些细胞进行标记。放射性分布与在标记起点的细胞中观察到的相似。相比之下,在允许温度下短暂标记的指数生长细胞显示出放射性的随机分布。这些数据表明复制起点和终点位于细胞极。