Kim Heesu, Noton Sarah L
Department of Virology, Immunology & Microbiology, Boston University Chobanian & Avedisian School of Medicine, Boston, MA, USA.
Methods Mol Biol. 2025;2948:109-126. doi: 10.1007/978-1-0716-4666-3_8.
The respiratory syncytial virus (RSV) minigenome system is a powerful analytical tool that has enabled viral transcription and genome replication to be modeled in a cellular context without viral infection. The minigenome assay can be used to dissect the different steps in RSV transcription and genome replication, assess the effects of mutations in cis-acting RNA elements, analyze mutations in the polymerase and other proteins in the transcription-replication machinery, and assess the effect of antiviral compounds on RSV polymerase functions. Here, we describe the RSV minigenome assay and detail a northern blotting technique for detecting viral mRNA transcripts and genome replication products.
呼吸道合胞病毒（RSV）微型基因组系统是一种强大的分析工具，它能够在无病毒感染的细胞环境中模拟病毒转录和基因组复制。微型基因组检测可用于剖析RSV转录和基因组复制的不同步骤，评估顺式作用RNA元件中突变的影响，分析转录-复制机制中聚合酶和其他蛋白质的突变，并评估抗病毒化合物对RSV聚合酶功能的影响。在此，我们描述了RSV微型基因组检测，并详细介绍了一种用于检测病毒mRNA转录本和基因组复制产物的Northern印迹技术。
