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Generation of Pseudorabies-resistant Animals by Germ-line Transformation

通过种系转化产生抗伪狂犬病动物

基本信息

批准号：
12556048
负责人：
ONO Etsuro
金额：
$ 7.3万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

项目摘要

To examine the activity of several gene promoters of pseudorabies virus (PRV) in vivo, we have generated transgenic mice expressing transgenes under the control of the viral promoters. Analyses of the tissue specificity of transgene expression demonstrated that the immediate-early gene promoter is neuronal tissue-specific, the early protein 0 gene promoter is a pan-specific and the latency associated transcript gene promoter is a neuron-specific, respectively.In order to assess the antiviral potential of PRV IE180 and a dominant-negative mutant of EP0 in vivo, several transgenic mouse lines expressing IE180 or the dominant-negative mutant of EP0 were generated. Experimental infection to these transgenic mice is now in progress.Herpesvirus entry mediator (HVEM) is a member of the tumor necrosis factor receptor family used as a cellular receptor by virion glycoprotein D (gD) of herpes simplex virus (HSV). In order to assess the antiviral potential of HVEM in vivo, three transgenic mouse lines expressing a soluble form of HVEM (HVEMlg) consisting of an extracellular domain of murine HVEM and the Fc portion of human lgG1 were generated. All of the transgenic mouse lines showed marked resistance to HSV-1 infection. To adapt this finding for generation of pseudorabies-resistant animal, we used porcine herpesvirus entry mediator C (HveC). Vero cells were transformed with the chimeric gene expressing a fusion protein consisting of an extracellular domain of porcine HveC and the Fc portion of human lgG1. The transformed cell lines expressing the fusion protein showed marked resistance to PRV infection. Generation of transgenic mice expressing the fusion protein is now in progress. These findings suggest the potential value of the fusion protein in agricultural livestock for enhanced disease resistance to pseudorabies.

为了研究伪狂犬病病毒（PRV）几种基因启动子在体内的活性，我们建立了在病毒启动子控制下表达转基因的转基因小鼠。对转基因表达的组织特异性分析表明，即早基因启动子具有神经组织特异性，早期蛋白0基因启动子具有泛特异性，潜伏相关转录基因启动子具有神经组织特异性。产生了几种表达IE 180或EP 0显性失活突变体的转基因小鼠系。疱疹病毒进入介体（HVEM）是单纯疱疹病毒（HSV）糖蛋白D（gD）的细胞受体，是肿瘤坏死因子受体家族的成员之一。为了评估HVEM在体内的抗病毒潜力，产生了表达由鼠HVEM的胞外结构域和人IgG 1的Fc部分组成的HVEM的可溶形式（HVEMlg）的三种转基因小鼠系。所有转基因小鼠品系均表现出对HSV-1感染的显著抗性。为了使这一发现适应伪狂犬病抗性动物的产生，我们使用了猪疱疹病毒进入介体C（HveC）。用表达由猪HveC的胞外结构域和人IgG 1的Fc部分组成的融合蛋白的嵌合基因转化Vero细胞。表达融合蛋白的转化细胞系对PRV感染表现出明显的抗性。表达融合蛋白的转基因小鼠的产生正在进行中。这些发现表明融合蛋白在农业家畜中用于增强对伪狂犬病的抗病性的潜在价值。