Pelchat M, Coté F, Perreault J P
Département de Biochimie, Faculté de Médecine, Université de Sherbooke, Canada.
Arch Virol. 2001;146(9):1753-63. doi: 10.1007/s007050170061.
We have developed an in vitro transcriptional assay using Escherichia coli RNA polymerase to initiate the replication of peach latent mosaic viroid (PLMVd). Regardless of the polarity of the PLMVd strand used as template, initiation in vitro occurred at the same hairpin structure. These initiation sites correspond to the 5'-ends of two small (280 nt) PLMVd-related RNAs found in infected peach leaves. Using a series of truncated PLMVd-derived transcripts, we have demonstrated that the viroid domain composed solely of the self-complementary hammerhead sequences is sufficient to trigger polymerase-driven replication in vitro. These data suggest that the bacterial-like RNA polymerase from peach chloroplasts catalyzes PLMVd replication.
我们开发了一种体外转录测定法，使用大肠杆菌RNA聚合酶来启动桃潜隐花叶类病毒（PLMVd）的复制。无论用作模板的PLMVd链的极性如何，体外起始都发生在相同的发夹结构处。这些起始位点对应于在受感染的桃叶中发现的两种小（280 nt）PLMVd相关RNA的5'末端。使用一系列截短的PLMVd衍生转录本，我们已经证明仅由自我互补锤头序列组成的类病毒结构域足以在体外触发聚合酶驱动的复制。这些数据表明，来自桃叶绿体的类细菌RNA聚合酶催化PLMVd复制。
