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Development of reverse genetics for Ibaraki virus to produce viable VP6-tagged IBAV

机译：茨城病毒逆向遗传学的发展，以产生可行的VP6标签IBAV

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Ibaraki virus (IBAV) is a member of the epizootic hemorrhagic disease virus (EHDV) serogroup, which belongs to the Orbivirus genus of the Reoviridae family. Although EHDV, including IBAV, represents an ongoing threat to livestock in the world, molecular mechanisms of EHDV replication and pathogenesis have been unclear. The reverse genetics (RG) system is one of the strong tools to understand molecular mechanisms of virus replication. Here, we developed a RG system for IBAV to identify the nonessential region of a minor structural protein, VP6, by generating VP6-truncated IBAV. Moreover, several tags were inserted into the truncated region to produce VP6-tagged IBAV. We demonstrated that all VP6-tagged IBAV could replicate in BHK cells in the absence of any helper VP6 protein. Further, tagged-VP6 proteins were first assembled into puncta in cells infected with VP6-tagged IBAV. Our data suggests that, in order to initiate primary replication, IBAV VP6 is likely to accumulate in some parts of infected cells to assemble efficiently into the primary replication complex (subcore).
机译：茨城病毒（IBAV）是流行性出血性疾病病毒（EHDV）血清群的成员，属于呼肠孤病毒科的Orbivirus属。尽管包括IBAV在内的EHDV代表着世界范围内对牲畜的持续威胁，但EHDV复制和发病机理的分子机制尚不清楚。反向遗传学（RG）系统是了解病毒复制的分子机制的强大工具之一。在这里，我们为IBAV开发了RG系统，以通过生成VP6截短的IBAV来鉴定次要结构蛋白VP6的非必需区域。此外，将几个标签插入到截短的区域中以产生VP6标签的IBAV。我们证明了在没有任何辅助VP6蛋白的情况下，所有带有VP6标签的IBAV均可在BHK细胞中复制。此外，标记的VP6蛋白首先在被VP6标记的IBAV感染的细胞中组装成点状。我们的数据表明，为了启动一级复制，IBAV VP6可能会在受感染细胞的某些部位积聚，从而有效地组装到一级复制复合体（子核心）中。

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