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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Replication of plant RNA virus genomes in a cell-free extract of evacuolated plant protoplasts
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Replication of plant RNA virus genomes in a cell-free extract of evacuolated plant protoplasts

机译：在无细胞植物原生质体的无细胞提取物中复制植物RNA病毒基因组

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摘要

The replication of eukaryotic positive-strand RNA virus genomes occurs through a complex process involving multiple viral and host proteins and intracellular membranes. Here we report a cell-free system that reproduces this process in vitro. This system uses a membrane-containing extract of uninfected plant protoplasts from which the vacuoles had been removed by Percoll gradient centrif-ugation. We demonstrate that the system supported translation, negative-strand RNA synthesis, genomic RNA replication, and sub-genomic RNA transcription of tomato mosaic virus and two other plant positive-strand RNA viruses. The RNA synthesis, which depended on translation of the genomic RNA, produced virus-related RNA species similar to those that are generated in vivo. This system will aid in the elucidation of the mechanisms of genome replication in these viruses.
机译：真核正链RNA病毒基因组的复制是通过一个复杂的过程进行的，该过程涉及多个病毒和宿主蛋白以及细胞内膜。在这里，我们报告了一个无细胞系统，该系统可在体外复制该过程。该系统使用未感染的植物原生质体的含膜提取物，通过Percoll梯度离心从中除去了液泡。我们证明该系统支持番茄花叶病毒和其他两种植物正链RNA病毒的翻译，负链RNA合成，基因组RNA复制和亚基因组RNA转录。 RNA合成依赖于基因组RNA的翻译，产生的病毒相关RNA种类与体内产生的相似。该系统将有助于阐明这些病毒中的基因组复制机制。

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