Project/Area Number |
10556073
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Research Category |
Grant-in-Aid for Scientific Research (B)
|
Allocation Type | Single-year Grants |
Section | 展開研究 |
Research Field |
Applied veterinary science
|
Research Institution | The University of Tokyo (2000-2001) Osaka Prefecture University (1998-1999) |
Principal Investigator |
HORIMOTO Taisuke The Institute of Medical Science, The University of Tokyo, Associate Professor, 医科学研究所, 助教授 (00222282)
|
Co-Investigator(Kenkyū-buntansha) |
MAEDA Ken Faculty of Agriculture, Yamaguchi University, Associate Professor, 農学部, 助教授 (90284273)
KAWAGUCHI Yasushi Medical Research Insutitute, Tokyo Medical and Dental University, Associate Professor, 難治疾患研究所, 助教授 (60292984)
SUGII Shunji Graduate School of Agriculture, Osaka Prefecture University, Professor, 大学院・農学生命科学研究科, 教授 (70162865)
TUCHIYA Kotaro Nippon Institute of Biological Science, Research Laboratory, Senior Researcher, 研究部, 主任研究員 (70207405)
GOTO Hideo The Institute of Medical Science, The University of Tokyo, Assitant Professor, 医科学研究所, 助手 (50323639)
田島 朋子 大阪府立大学, 農学部, 助手 (90173145)
|
Project Period (FY) |
1998 – 2001
|
Project Status |
Completed (Fiscal Year 2001)
|
Budget Amount *help |
¥12,300,000 (Direct Cost: ¥12,300,000)
Fiscal Year 2001: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2000: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1999: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1998: ¥3,900,000 (Direct Cost: ¥3,900,000)
|
Keywords | Porcine cytomegalovirus / Betaherpesvirus / Viral vector / DNA polymerase / Capsid protein / Glycoprotein / Vaccine / PCR / 膜糖蛋白 / カプシッド蛋白 |
Research Abstract |
To develop a recombinant viral live vaccine for pigs, we have to select vector virus. In this project, we sought to use porcine cytomegalovirus (PCMV) as a vector virus, because of its low pathogenicity for pigs, its capacity in accommodating foreign antigens and its persistent infectious property leading to long lasting antigen presentation. To this end, we initially have to gain virological data of PCMV genome and protein organization and structure. During this research period, we have achieved as follows: (1) Resrtiction endonuclease fragments of PCMV genome DNA were cloned and randomly sequenced to search for PCMV genes. (2) Nucleotide sequences of major genes essential for viral replication, such as DNA polymerase, major capsid protein (MCP), glycoprotein B (gB) genes, and their flanking gene clusters, were determined. Protein structure was analyzed by predicted amino acid sequences. (3) Phylogenetic analysis using these PCMV genes sequences established that PCMV is a betaherpesvirus closely related to human herpesvirus 6 and 7, suggesting that PCMV system may provide a promising animal model for these two human herpesviruses. (4) We successfully expressed MCP and its deletion derivatives. (5) PCR protocols based on MCP gene sequences were established and applied to blood-filter paper samples. These results provide basic information required for establishment of PCMV vector system used for production of a recombinant viral live vaccine for pigs. Also these data may contribute to basic study for xenotransplantation from pigs to human in the future. Bacterial artificial chromosomal system has been established for some herpesvirus genome DNA manipulation such as herpes simplex virus, indicating that recombinant PCMV should be achieved in the near future, leading to production of a recombinant viral live vaccine for pigs.
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