| Project/Area Number |
07556069
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Applied veterinary science
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| Research Institution | The University of Tokyo |
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Principal Investigator |
MIKAMI Takeshi Univ.of Tokyo, Grad.Sch.of Agri.and Life Sci., Professor, 大学院・農学生命科学研究科, 教授 (20091506)
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| Co-Investigator(Kenkyū-buntansha) |
MAEDA Ken (OTSUKA Haruki) Yamaguchi University, Faculty of Agriculture, Assosiate Professor, 農学部, 助教授 (80261957)
HORIMOTO Taisuke Osaka Prefecture University, Faculty of Agriculture, Associate Professor, 農学部, 助教授 (00222282)
MIYAZAWA Takayuki Univ.of Tokyo, Grad.Sch.of Agri.and Life Sci.Assistant Professor, 大学院・農学生命科学研究科, 助手 (80282705)
TSUJIMOTO Hajime Univ.of Tokyo, Grad.Sch.of Agri.and Life Sci., Associate Professor, 大学院・農学生命科学研究科, 助教授 (60163804)
前田 健 山口大学, 農学部, 助教授 (90284273)
遠矢 幸伸 鹿児島大学, 農学部, 助教授 (30021702)
望月 雅美 共立商事(株), 臨床微生物研究所, 所長(研究職) (90157834)
時吉 幸男 化学及血清療法研究所, 研究開発部, 室長(研究職)
藤川 勇治 共立商事(株), 中央研究所, 副所長(研究職)
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| Project Period (FY) |
1995 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥19,700,000 (Direct Cost: ¥19,700,000)
Fiscal Year 1997: ¥5,400,000 (Direct Cost: ¥5,400,000)
Fiscal Year 1996: ¥5,700,000 (Direct Cost: ¥5,700,000)
Fiscal Year 1995: ¥8,600,000 (Direct Cost: ¥8,600,000)
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| Keywords | recombinant vaccine / feline herpesvirus 1 / feline calicivirus / feline immunodeficiency virus / gag protein / capsid protein / thymidine kinase gene / immunoblot analysis / FHV-1 / TK / FCV / 間接蛍光抗体法 / イムノブロット解析 / FIV / 組換え多価ワクチン / ウイルス性鼻気管炎 / 組換えウイルス / TK遺伝子欠損株 / 伝染性鼻気管炎 / 遺伝子マーカー |
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| Research Abstract |
The aim of the present studies is to develop polyvalent recombinant (rec) vaccines which are superior to safeness, inexpensiveness and effctiveness to cats, and to put the vaccines to practical use. The following results are obtained.
1.We succeeded in the attenuation of thymidine kinase (TK) deficient mutant (C7301dlTK) of feline herpesvirus type 1 (FHV-1) in cats and the construction of a recombinat FHV-1 (C7301dlTK-Cap) inserted a precursor capsid gene of feline calicivirus (FCV) into the TK deletion locus of the C7301dlTK.Cats vaccinated with C7301dlTK-Cap and then challenged with virulent FHV-1 and FCV were protected to a significant degree against the manifestations of the illness caused by both viruses. However, immune presponse to FCV was weak.
2.Therfore to improve further on the recombinat FHV-1, we constructed an improved recombinant FHV-1 (dlTK (gCp) -Cap) carring a putative FHV-1 gC promoter sequence on the upstream of the FCV precursor capsid gene of the C7301dlTK-Cap. Growth kinetics of the dlTK (gCp) -Cap in cell cultures was similar to those of C7301dlTK and C7301dlTK-Cap. A strong expression of FCV immunogenic antigen by dlTK (gCp) -Cap was confirmed by indirect immunofluorescence and enzime-linked immunosorbent assays. In addition, one vaccination with dlTK (gCp) -Cap protected cats more effectively against subsequent virulent FCV challenge than that with C7301dlTK-Cap.
3.We constructed a recombinat FHV-1 expressing Gag protein of feline immunodeficiency virus (FIV), in which a cDNA encoding the Gag protein of FIV was inserted at the TK deletion site of C7301ddlTK,and was designated as C7301ddlTK-gag. Growth kinetics of the reconbinat viruses in Crandell feline kidney cells was similar to that of the parent C7301 strain. By immunoblot analysis, C7301ddlTK-gag was confirmed to express the FIV Gag precursor protein in the cells.
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