噬菌体φX174复制型DNA的不连续复制
Discontinuous replication of replicative form DNA from bacteriophage phiX174.
作者信息
Machida Y, Okazaki T, Okazaki R
出版信息
Proc Natl Acad Sci U S A. 1977 Jul;74(7):2776-9. doi: 10.1073/pnas.74.7.2776.
Abstract
Bacteriophage phiX174 DNA has been labeled with short pulses of [3H]thymidine during synthesis of replicative form molecules in infected Escherichia coli HF4704 cells. The replicating phiX174 DNA was isolated and analyzed by sedimentation in an alkaline sucrose gradient. During a brief pulse (5 sec at 30 degrees), the radioactivity incorporated into the complementary strand was found in chains much shorter than one genome length. Of the radioactivity incorporated into the viral strand, two-thirds was in the short pieces and the rest was in chains of one genome length or longer. RNA attachment to the 5' end of both strand components of the nascent short pieces was shown by the appearance of spleen exonuclease-digestable nascent molecules after alkali treatment. These observations suggest that the viral as well as the complementary strand is synthesized by the discontinuous mechanism with RNA primers during replication of duplex phiX174 DNA.
摘要
在感染的大肠杆菌HF4704细胞中合成复制型分子期间，用短脉冲的[3H]胸苷标记了噬菌体phiX174 DNA。分离出正在复制的phiX174 DNA，并通过在碱性蔗糖梯度中沉降进行分析。在短暂脉冲（30℃下5秒）期间，发现掺入互补链中的放射性存在于比一个基因组长度短得多的链中。在掺入病毒链的放射性中，三分之二存在于短片段中，其余存在于一个基因组长度或更长的链中。碱处理后出现脾脏外切核酸酶可消化的新生分子，表明RNA附着于新生短片段的两条链成分的5'端。这些观察结果表明，在双链phiX174 DNA复制期间，病毒链和互补链都是通过RNA引物的不连续机制合成的。
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