| Project/Area Number |
05506002
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
Basic veterinary science/Basic zootechnical science
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
KIDA Hiroshi (1994-1995) Hokkaido Univ., Grad. Sch. of Vet. Med., Pro., 大学院獣医学研究科, 教授 (10109506)
清水 悠紀臣 (1993) 北海道大学, 獣医学部, 教授 (80206218)
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| Co-Investigator(Kenkyū-buntansha) |
OKABE Tatsuji Kyoto Biken Lab., Manager, 部長
SEKIKAWA Kenji Nation. Insti. on Ani. Health, The head of anoffice, 家畜衛生試験場, 室長
MIKAMI Takeshi Tokyo Univ., Fac. of Agri., Pro., 農学部, 教授 (20091506)
KASAI Noriyuki Tohoku Univ., Fac. of Med., Pro., 医学部, 教授 (60001947)
ONO Etsuro Hokkaido Univ., Insti. fo Immunol. Sci., Associ. Pro., 免疫化学研究所, 助教授 (00160903)
大塚 治城 東京大学, 農学部, 教授
喜田 宏 北海道大学, 獣医学部, 助教授 (10109506)
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| Project Period (FY) |
1993 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥29,900,000 (Direct Cost: ¥29,900,000)
Fiscal Year 1995: ¥4,900,000 (Direct Cost: ¥4,900,000)
Fiscal Year 1994: ¥9,000,000 (Direct Cost: ¥9,000,000)
Fiscal Year 1993: ¥16,000,000 (Direct Cost: ¥16,000,000)
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| Keywords | Aujeszky's disease virus / Herpesvirus / Transcription / Transcriptional repression / ヘルペスウイルス / オーエスキー病 |
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| Research Abstract |
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.
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