Okuno Y, Okazaki T, Masukata H
Department of Molecular Biology, School of Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 564-01, Japan.
Nucleic Acids Res. 1997 Feb 1;25(3):530-7. doi: 10.1093/nar/25.3.530.
We have identified five autonomously replicating sequences (ARSs) in a 100 kbp region of the Schizosaccharomyces pombe chromosome II. Analyses of replicative intermediates of the chromosome DNA by neutral/neutral two-dimensional gel electrophoresis demonstrated that at least three of these ARS loci operate as chromosomal replication origins. One of the loci,ori2004, was utilized in almost every cell cycle, while the others were used less frequently. The frequency of initiation from the respective chromosomal replication origin was found to be roughly proportional to the efficiency of autonomous replication of the corresponding ARS plasmid. Replication from ori2004 was initiated within a distinct region almost the same as that for replication of the ARS plasmid. These results showed that the ori2004 region of approximately 3 kbp contains all the cis elements essential for initiation of chromosome replication.
我们在粟酒裂殖酵母二号染色体的100 kbp区域内鉴定出了五个自主复制序列（ARSs）。通过中性/中性二维凝胶电泳对染色体DNA复制中间体进行分析表明，这些ARS位点中至少有三个作为染色体复制起点发挥作用。其中一个位点ori2004在几乎每个细胞周期中都被利用，而其他位点的使用频率较低。发现从各个染色体复制起点起始的频率大致与相应ARS质粒的自主复制效率成正比。ori2004的复制在一个与ARS质粒复制几乎相同的特定区域内起始。这些结果表明，大约3 kbp的ori2004区域包含了染色体复制起始所必需的所有顺式元件。
