Molecular pathogenic study on neuropathogenicity of a new lethal and neurotropic herpesvirus

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Equine herpesvirus type 9 (EHV-9) is a new herpesvirus isolated from brains of Thomson's gazelles affected by encephalitis in a zoo. EHV-9 showed strong neurovirulence and a broad host spectrum. I would like to find the molecular mechanism of the strong neurotropism and neurovirulence of EHV-9. I tried to construct a bacterial artificial chromosome clone of EHV-9 genome in Escherichi coli in this year.At first, an expression marker of a green fluorescence protein gene (GFP) was instered in the non-coding region of EHV-9 genome. The cassette of GFP gene contained loxP sequences at the each end of the gene. The insertion point was at the middle of ORF62 and ORF63. The virus inserted GFP was referenced as EHV-9-GFP. The GFP cassette was deleted from EHV-9-GFP using a Cre recombinase. The virus deleted GFP contained one copy of loxP in its genome. The virus was EHV-9-loxP. I tried to insert pBeloBAC plasmid containing one copy of loxP into EHV-9-loxP by the Cre recombinase. At the time of this report, I could not obtain any clone of EHV-9 inserted pBeloBAC. I am testing various conditions for obtaining EHV-9 instered pBeloBAC.