Bataille D, Epstein A
Centre de Génétique Moléculaire et Cellulaire, UMR 106-CNRS, Université Claude Bernard Lyon I, Villeurbanne, France.
Virology. 1994 Sep;203(2):384-8. doi: 10.1006/viro.1994.1498.
The structure of herpes simplex virus (HSV-1) replicative DNA was studied employing pulse-field gel electrophoresis to allow separation and direct visualization of large size virus DNA fragments. Accumulation of HSV-1 DNA that fails to enter the gel was detected from 6 hr postinfection. This corresponds, at least in part, to high molecular weight replicative intermediates. Analysis of this DNA employing restriction enzymes SpeI and AseI, which cut only once and twice respectively in the HSV-1 genome, revealed that these intermediates are concatemeric and contain adjacent genomes with L components in different orientations. Adjacent genomes with L components in the same orientation or with L components in inverted orientations were found in similar amounts, suggesting that inversions occur frequently. Our results suggest that inversions of L components are generated by a quite efficient mechanism, perhaps during replication. Observations made in this work could be explained by rolling circle-based models of replication of HSV-1 genomes, with replication starting at the different origins and/or combined with strand transfer at inverted repeats.
利用脉冲场凝胶电泳技术研究单纯疱疹病毒1型（HSV - 1）复制性DNA的结构，以实现对大尺寸病毒DNA片段的分离和直接可视化。从感染后6小时开始检测到未进入凝胶的HSV - 1 DNA的积累。这至少部分对应于高分子量复制中间体。使用限制性内切酶SpeI和AseI对该DNA进行分析，它们在HSV - 1基因组中分别仅切割一次和两次，结果显示这些中间体是串联体，并且包含具有不同方向L成分的相邻基因组。发现具有相同方向L成分或反向L成分的相邻基因组数量相似，这表明倒位频繁发生。我们的结果表明，L成分的倒位是由一种相当有效的机制产生的，可能发生在复制过程中。这项工作中的观察结果可以通过基于滚环的HSV - 1基因组复制模型来解释，复制从不同的起始点开始和/或与反向重复序列处的链转移相结合。
