Braunstein J D, Schildkraut C L
Biochem Biophys Res Commun. 1984 Aug 30;123(1):108-13. doi: 10.1016/0006-291x(84)90386-3.
The time of replication in S phase was determined for the 7.3 kb EcoRI segment containing the beta-major globin gene and the 14 kb EcoRI segment containing the beta-minor globin gene in a murine Friend erythroleukemia virus transformed (MEL) cell line. Cells were obtained from different intervals of S phase by centrifugal elutriation to avoid artifacts of chemical synchronization. Newly synthesized DNA from different parts of S phase were obtained by isolation of 5-bromouracil (BU) labelled DNA from these cells. The BU-DNA synthesized during four different intervals of S was transferred to diazobenzyloxymethyl (DBM) paper and hybridized with a beta-globin cDNA probe. Quantitation showed that both beta-globin segments were replicated in the first quarter of the S phase with no significant difference in their time of replication in this MEL cell line.
在一个鼠类Friend红白血病病毒转化(MEL)细胞系中,测定了含有β-珠蛋白基因的7.3 kb EcoRI片段和含有β-珠蛋白基因的14 kb EcoRI片段在S期的复制时间。通过离心淘洗从S期的不同时间间隔获取细胞,以避免化学同步化带来的假象。通过从这些细胞中分离5-溴尿嘧啶(BU)标记的DNA,获得了来自S期不同部分新合成的DNA。在S期四个不同时间间隔合成的BU-DNA被转移到重氮苄氧基甲基(DBM)纸上,并与β-珠蛋白cDNA探针杂交。定量分析表明,在这个MEL细胞系中,两个β-珠蛋白片段均在S期的第一个四分之一时间段进行复制,它们的复制时间没有显著差异。