Department of Infectious and Parasitic Diseases, Virology and Viral Diseases, Faculty of Veterinary Medicine, University of Liège, 4000 Liège, Belgium.
J Gen Virol. 2010 Nov;91(Pt 11):2723-33. doi: 10.1099/vir.0.024109-0. Epub 2010 Aug 11.
Based on sequencing data, norovirus (NoV) recombinants have been described, but no experimental evidence of recombination in NoVs has been documented. Using the murine norovirus (MNV) model, we investigated the occurrence of genetic recombination between two co-infecting wild-type MNV isolates in RAW cells. The design of a PCR-based genotyping tool allowed accurate discrimination between the parental genomes and the detection of a viable recombinant MNV (Rec MNV) in the progeny viruses. Genetic analysis of Rec MNV identified a homologous-recombination event located at the ORF1-ORF2 overlap. Rec MNV exhibited distinct growth curves and produced smaller plaques than the wild-type MNV in RAW cells. Here, we demonstrate experimentally that MNV undergoes homologous recombination at the previously described recombination hot spot for NoVs, suggesting that the MNV model might be suitable for in vitro studies of NoV recombination. Moreover, the results show that exchange of genetic material between NoVs can generate viruses with distinct biological properties from the parental viruses.
基于测序数据,已描述了诺如病毒(NoV)重组体,但尚未记录到 NoV 中重组的实验证据。我们使用鼠诺如病毒(MNV)模型,研究了两种共感染的野生型 MNV 分离株在 RAW 细胞中发生遗传重组的情况。基于 PCR 的基因分型工具的设计允许在亲代基因组之间进行准确区分,并在子代病毒中检测到有活力的重组 MNV(Rec MNV)。对 Rec MNV 的遗传分析鉴定了位于 ORF1-ORF2 重叠处的同源重组事件。Rec MNV 在 RAW 细胞中的生长曲线和产生的蚀斑均明显小于野生型 MNV。在这里,我们通过实验证明 MNV 在先前描述的 NoV 重组热点处发生了同源重组,这表明 MNV 模型可能适合用于 NoV 重组的体外研究。此外,结果表明,NoV 之间遗传物质的交换可以产生与亲代病毒具有不同生物学特性的病毒。