title: Development of reverse genetics for Ibaraki virus to produce viable VP6-tagged IBAV.
abstract: 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).
authors.affiliation: Microbiology & Immunology, Division of Animal Science, Department of Bioresource Science, Graduate School of Agricultural Science, Kobe University, 1-1, Rokkodai, Nada-ku, Kobe-city 657-8501, Japan.
authors.forename: Eiko
authors.initials: E
authors.lastname: Matsuo
authors.affiliation: Microbiology & Immunology, Division of Animal Science, Department of Bioresource Science, Graduate School of Agricultural Science, Kobe University, 1-1, Rokkodai, Nada-ku, Kobe-city 657-8501, Japan.
authors.forename: Keiichi
authors.initials: K
authors.lastname: Saeki
authors.affiliation: Department of Pathogen Molecular Biology, Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT, UK.
authors.forename: Polly
authors.initials: P
authors.lastname: Roy
authors.affiliation: Microbiology & Immunology, Division of Animal Science, Department of Bioresource Science, Graduate School of Agricultural Science, Kobe University, 1-1, Rokkodai, Nada-ku, Kobe-city 657-8501, Japan.
authors.forename: Junichi
authors.initials: J
authors.lastname: Kawano
keywords: EHD, epizootic hemorrhagic disease; EHDV, epizootic hemorrhagic disease virus; Epizootic hemorrhagic disease virus; IBAV, Ibaraki virus; Ibaraki virus; Primary replication; RG, reverse genetics; Reverse genetics system; VIB, virus inclusion body; VP6; Viable VP6-tagged virus