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

马疱疹病毒9型（EHV-9）是从动物园感染脑炎的汤氏瞪羚脑组织中分离到的一种新型疱疹病毒。EHV-9具有较强的神经毒力和广泛的宿主谱。我希望找到EHV-9强嗜神经性和神经毒力的分子机制。本研究尝试在大肠杆菌中构建EHV-9基因组的细菌人工染色体克隆，首先在EHV-9基因组的非编码区插入一个绿色荧光蛋白基因（GFP）的表达标记。GFP基因的盒在基因的每一端含有loxP序列。插入点位于ORF 62和ORF 63的中间。将插入GFP的病毒称为EHV-9-GFP。使用Cre重组酶从EHV-9-GFP中删除GFP盒。缺失GFP的病毒在其基因组中含有一个loxP拷贝。该病毒为EHV-9-loxP。我尝试通过Cre重组酶将含有一个拷贝的loxP的pBeloBAC质粒插入EHV-9-loxP中。在撰写本报告时，我无法获得插入EHV-9的pBeloBAC的任何克隆。我正在测试获得EHV-9 instered pBeloBAC的各种条件。

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.