Kaufmann G, Bar-Shavit R, DePamphilis M L
Nucleic Acids Res. 1978 Jul;5(7):2535-45. doi: 10.1093/nar/5.7.2535.
Simian virus 40 replicating DNA was pulse labeled with alpha-32P-dATP using an acellular DNA replication system. Nascent DNA chains of less than 200 nucleotides (Okazaki pieces) were then isolated from the denatured replicating DNA by electrosieving through a polyacrylamide gel column. The purified Okazaki pieces were hybridized to separated strands of Bg1(1)+Hpa1 simian virus 40 DNA restriction fragments immobilized on nitrocellulose filters. Only strands with polarity of the DNA replication fork direction hybridized with Okazaki pieces. Hence, Okazaki pieces in simian virus 40 are synthesized against the DNA replication fork direction.
使用无细胞DNA复制系统，用α-32P-dATP对猴病毒40复制DNA进行脉冲标记。然后通过聚丙烯酰胺凝胶柱电筛从变性的复制DNA中分离出小于200个核苷酸的新生DNA链（冈崎片段）。将纯化的冈崎片段与固定在硝酸纤维素滤膜上的Bg1(1)+Hpa1猴病毒40 DNA限制片段的单链进行杂交。只有具有DNA复制叉方向极性的链与冈崎片段杂交。因此，猴病毒40中的冈崎片段是逆着DNA复制叉方向合成的。
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