Huberman J A, Spotila L D, Nawotka K A, el-Assouli S M, Davis L R
Department of Molecular and Cellular Biology, Roswell Park Memorial Institute, Buffalo, New York 14263.
Cell. 1987 Nov 6;51(3):473-81. doi: 10.1016/0092-8674(87)90643-x.
We have used two-dimensional neutral/alkaline agarose gel electrophoresis to separate the nascent strands of replicating yeast 2 micron plasmid DNA molecules according to extent of replication, away from nonreplicating molecules and parental strands. Analysis of the lengths of nascent strands by sequential hybridization with short probes shows that replication proceeds bidirectionally from a single origin at map position 3700 +/- 100, coincident with the genetically mapped ARS element. The two recombinational isomers of 2 microns plasmid (forms A and B) replicate with equal efficiency. These results suggest that ARS elements may prove to be replication origins for chromosomal DNA.
我们使用二维中性/碱性琼脂糖凝胶电泳，根据复制程度将正在复制的酵母2微米质粒DNA分子的新生链与非复制分子及亲代链分离开来。通过与短探针进行连续杂交来分析新生链的长度，结果表明复制从图谱位置3700±100处的单个起点双向进行，这与基因定位的自主复制序列（ARS）元件一致。2微米质粒的两种重组异构体（A和B型）复制效率相同。这些结果表明，ARS元件可能被证明是染色体DNA的复制起点。
