Centro de Biotecnología y Genómica de Plantas, Universidad Politécnica de Madrid (UPM)-Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA/CSIC), Madrid, Spain.
Departamento de Genética, Facultad de Biología, Universidad de Sevilla, Sevilla, Spain.
Methods Mol Biol. 2024;2751:47-68. doi: 10.1007/978-1-0716-3617-6_4.
The most important advances in our understanding of the viral life cycle, such as genome replication, packaging, transmission, and host interactions, have been made via the development of viral infectious full-length clones. Here, we describe the detailed protocols for the construction of an infectious clone derived from Botrytis virus F (BVF), a mycoflexivirus infecting the plant pathogenic fungus Botrytis cinerea, the determination of the complete sequence of the cloned mycovirus, the preparation of fungal protoplasts, and the transfection of protoplasts using transcripts derived from the BVF infectious clone.
我们对病毒生命周期的理解最重要的进展，如基因组复制、包装、传播和宿主相互作用，都是通过开发病毒传染性全长克隆实现的。在这里，我们描述了从感染植物病原菌 Botrytis cinerea 的真菌弹状病毒 Botrytis 病毒 F（BVF）衍生的传染性克隆的构建、克隆真菌病毒的全长序列的确定、真菌原生质体的制备以及使用源自 BVF 传染性克隆的转录本转染原生质体的详细方案。
