A new approach for the control of Aujeszky's disease by using germ-line transformation
利用种系转化控制 Aujeszky 病的新方法
基本信息
- 批准号:05506002
- 负责人:
- 金额:$ 19.14万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Developmental Scientific Research (A)
- 财政年份:1993
- 资助国家:日本
- 起止时间:1993 至 1995
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Intracellular immunization using dominant-negative mutants of viral proteins is prorposed as a new approach to antiviral therapy in humans and to germ-line transformation in animals to confer resistance to viral infections. To obtain effective repressors of Aujeszky's disease virus (ADV) immediate-early (IE) gene expression, mutant genes encoding dominant-negative mutants of ADV IE protein (IE180) and early protein 0 (EPO), and a chimeric gene encoding a fusion protein consisting of the DNA-binding domain of IE180 and a tail-trucated herpes simplex virus 1 VP16 lacking the transcription activation domain were constructed. These gene products inhibited transcription from the ADV IE promoter in transient expression assays. HeLa cell lines stably transformed with the genes showed resistance to ADV infection. Among them, resistance of a cell line transformed with the chimeric transgene was remarkable.In order to assess the antiviral potential of the fusion protein in vivo, the chimeric gene under the control of polypepetide chain elongation factor 1alpha or the mouse Mx1 promoter was used for generation of transgenic mice. C57BL/6 zygotes were microinjected with about 1000 copies of the DNA fragment encoding the chimeric gene. Of the resulting 41 births, five animals had the transgene as determined by Southern blot analysis of tail DNA.Four of the founders failed to transmit the transgene to their progeny. A remaining line transmitted the transgene to F1 progeny. In the transgenic mice, the fusion protein is expressed under the control of the Mx1 promoter which is inducible by double stranded RNA,interferon, or virus infection. To assess the resistance of the established transgenic mice to ADV challenge, breeding of the F1 mice is now in progress.
使用病毒蛋白显性-负性突变体的细胞内免疫被认为是一种新的人类抗病毒治疗和动物生殖系转化的方法,以增强对病毒感染的抵抗力。为了获得有效抑制Aujeszky病病毒(ADV)即刻早期(IE)基因表达的基因,构建了编码ADV IE蛋白(IE180)和早期蛋白0(EPO)显性-阴性突变体的突变基因,以及编码IE180 DNA结合域与尾部结构的单纯疱疹病毒1型VP16融合蛋白的嵌合基因。在瞬时表达实验中,这些基因产物抑制了ADV IE启动子的转录。稳定转化的HeLa细胞株对ADV感染表现出抗性。为了评价融合蛋白在体内的抗病毒能力,将多肽链延长因子1α或小鼠Mx1启动子控制的嵌合基因用于转基因小鼠的产生。向C57BL/6受精卵显微注射约1000个拷贝的编码嵌合基因的DNA片段。通过对尾部DNA的Southern杂交分析,在得到的41只幼崽中,有5只动物具有转基因。其中4只动物未能将转基因传递给他们的后代。剩下的一株系将转基因传递给F1后代。在转基因小鼠中,融合蛋白在Mx1启动子的控制下表达,Mx1启动子可被双链RNA、干扰素或病毒感染诱导。为了评估已建立的转基因小鼠对ADV攻击的抵抗力,目前正在进行F1小鼠的培育。
项目成果
期刊论文数量(39)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Kawaguchi, Y.: "The feline herpesvirus type 1 ICP4 down-regulates feline immunodeficiency virus long terminal repeat (LTR) -directed gene expression via the C/EBP site in the LTR." Journal of Veterinary Medical Science. 57. 1129-1131 (1995)
Kawaguchi, Y.:“猫疱疹病毒 1 型 ICP4 通过 LTR 中的 C/EBP 位点下调猫免疫缺陷病毒长末端重复 (LTR) 定向基因表达。”
- DOI:
- 发表时间:
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- 影响因子:0
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- 通讯作者:
Kawaguchi, Y.: "The feline herpesvirus type 1 ICP4 down-reguates feline immunodeficiency virus long terminal repeat (LTR)-directed gene expression via the C/EBP site in the LTR." Journal of Veterinary Medical Science. 57 (6). 1129-1131 (1995)
Kawaguchi, Y.:“猫疱疹病毒 1 型 ICP4 通过 LTR 中的 C/EBP 位点下调猫免疫缺陷病毒长末端重复 (LTR) 定向基因表达。”
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- 发表时间:
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- 影响因子:0
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- 通讯作者:
Kawaguchi, Y.: "Molecular interaction between retroviruses and herpesviruses." Journal of Veterinary Medical Science. 57. 801-811 (1995)
Kawaguchi, Y.:“逆转录病毒和疱疹病毒之间的分子相互作用。”
- DOI:
- 发表时间:
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- 影响因子:0
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- 通讯作者:
Taharaguchi, S.: "Mapping of a functional region conferring nuclear localization of pseudorabies virus immediate-early protein." Archives of Virology. 140. 1737-1746 (1995)
Taharaguchi, S.:“对伪狂犬病病毒立即早期蛋白核定位的功能区域进行绘图。”
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- 期刊:
- 影响因子:0
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- 通讯作者:
Taharaguchi, S.: "Mapping of transcriptional regulatory domains of pseudorabies virus immediate-early protein." Archives of Virology. 137 (3-4). 289-303 (1994)
Taharaguchi, S.:“伪狂犬病病毒立即早期蛋白转录调控域的定位。”
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- 影响因子:0
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{{ truncateString('KIDA Hiroshi', 18)}}的其他基金
The role of Interleukin-6 family cytokines in repair and tumorigenesis in the lung.
Interleukin-6 家族细胞因子在肺部修复和肿瘤发生中的作用。
- 批准号:
20890116 - 财政年份:2008
- 资助金额:
$ 19.14万 - 项目类别:
Grant-in-Aid for Young Scientists (Start-up)
Ecology and pandemic planning of animal influenza virus
动物流感病毒的生态学和流行病规划
- 批准号:
15108004 - 财政年份:2003
- 资助金额:
$ 19.14万 - 项目类别:
Grant-in-Aid for Scientific Research (S)
Ecological study of animal influenza viruses : To prepare for the emergence of pandemic influenza
动物流感病毒的生态学研究:为大流行性流感的出现做好准备
- 批准号:
12375006 - 财政年份:2000
- 资助金额:
$ 19.14万 - 项目类别:
Grant-in-Aid for Scientific Research (A)
Molecular mechanisms of infection and pathogenesis of viruses
病毒感染和发病的分子机制
- 批准号:
10041151 - 财政年份:1998
- 资助金额:
$ 19.14万 - 项目类别:
Grant-in-Aid for Scientific Research (A).
Molecular mechanisms of infection and pathogenesis of viruses
病毒感染和发病的分子机制
- 批准号:
08406020 - 财政年份:1996
- 资助金额:
$ 19.14万 - 项目类别:
Grant-in-Aid for Scientific Research (A)
Prediction of new pandemic influenza viruses -Distribution of the viruses in waterfowls in their nesting places in Siberia
新型大流行性流感病毒的预测-西伯利亚筑巢地水禽中病毒的分布
- 批准号:
07041118 - 财政年份:1995
- 资助金额:
$ 19.14万 - 项目类别:
Grant-in-Aid for international Scientific Research
Origin and Evolution of Influenza Viruses
流感病毒的起源和进化
- 批准号:
05044117 - 财政年份:1993
- 资助金额:
$ 19.14万 - 项目类别:
Grant-in-Aid for international Scientific Research
Evolution and Prediction of Influenza Viruses
流感病毒的进化和预测
- 批准号:
03041013 - 财政年份:1991
- 资助金额:
$ 19.14万 - 项目类别:
Grant-in-Aid for international Scientific Research
Prediction and Control of New Pandemic Influenza Viruses
新型大流行性流感病毒的预测与控制
- 批准号:
01480103 - 财政年份:1989
- 资助金额:
$ 19.14万 - 项目类别:
Grant-in-Aid for General Scientific Research (B)
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