Price B D, Rueckert R R, Ahlquist P
Institute for Molecular Virology, University of Wisconsin, Madison 53706-1596, USA.
Proc Natl Acad Sci U S A. 1996 Sep 3;93(18):9465-70. doi: 10.1073/pnas.93.18.9465.
Here we describe the first instances to our knowledge of animal virus genome replication, and of de novo synthesis of infectious virions by a nonendogenous virus, in the yeast Saccharomyces cerevisiae, whose versatile genetics offers significant advantages for studying viral replication and virus-host interactions. Flock house virus (FHV) is the most extensively studied member of the Nodaviridae family of (+) strand RNA animal viruses. Transfection of yeast with FHV genomic RNA induced viral RNA replication, transcription, and assembly of infectious virions. Genome replication and virus synthesis were robust: all replicating FHV RNA species were readily detected in yeast by Northern blot analysis and yields of virions per cell were similar to those from Drosophila cells. We also describe in vivo expression and maintenance of a selectable yeast marker gene from an engineered FHV RNA derivative dependent on FHV-directed RNA replication. Use of these approaches with FHV and their possible extension to other viruses should facilitate identification and characterization of host factors required for genomic replication, gene expression, and virion assembly.
在此，据我们所知，我们描述了动物病毒基因组在酵母酿酒酵母中复制的首个实例，以及非内源性病毒从头合成感染性病毒粒子的首个实例，酵母具有多样的遗传学特性，为研究病毒复制和病毒-宿主相互作用提供了显著优势。禽传染性脑脊髓炎病毒（FHV）是正义链RNA动物病毒诺达病毒科中研究最为广泛的成员。用FHV基因组RNA转染酵母可诱导病毒RNA复制、转录以及感染性病毒粒子的组装。基因组复制和病毒合成十分活跃：通过Northern印迹分析在酵母中很容易检测到所有正在复制的FHV RNA种类，每个细胞产生的病毒粒子产量与果蝇细胞中的产量相似。我们还描述了来自工程化FHV RNA衍生物的可选择酵母标记基因在体内的表达和维持，该表达和维持依赖于FHV指导的RNA复制。将这些方法应用于FHV以及可能将其扩展到其他病毒，应有助于鉴定和表征基因组复制、基因表达和病毒粒子组装所需的宿主因子。
