2003 Fiscal Year Final Research Report Summary
Producing and Certification of Recombinant Antigen with LCMV Isolated in Japan for Diagnosis of LCMV Infection in Mice.
| Project/Area Number |
14580800
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Laboratory animal science
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| Research Institution | Nagasaki University |
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Principal Investigator |
OHSAWA Kazutaka Nagasaki University, Center for Frontier Life Sciences, Division of Comparative Medicine, Associate Professor, 先導生命科学研究支援センター, 助教授 (90244756)
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| Co-Investigator(Kenkyū-buntansha) |
SATO Hiroshi Nagasaki University, Center for Frontier Life Sciences, Division of Comparative Medicine, Professor, 先導生命科学研究支援センター, 教授 (50072947)
KATAMINE Shigeru Nagasaki University, Graduate School of Biomedical Sciences, Professor, 大学院・医歯薬学総合研究科, 教授 (40161062)
KONDO Takahito Nagasaki University, Graduate School of Biomedical Sciences, Professor, 大学院・医歯薬学総合研究科, 教授 (00158908)
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| Project Period (FY) |
2002 – 2003
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| Keywords | LCMV / Recombinant antigen |
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| Research Abstract |
In 1996, six different LCMV strains (OQ28,OQ32,OQ38,OQ48,OQ49 & OQ52) were isolated from wild mice of a port in Japan. Nucleotide sequence analysis of short (S) fragment, including Np and Gp gene, of these isolates showed: 1) high homology each others (>99%); 2) low similarity with prototype WE (84%) or Armstrong (85%) strains. Therefore we planed that should produce authentic recombinant antigens based on the sequence of isolated in Japan. Rapid Translation System (RTS) of E coli lysate and mammalian cell expression system were used for full Np gene and Gp regions, respectively. By RTS, full Np gene on vector pIVEX2.3 & pIVEX2.4b induced a protein of 65.9kDa, which fitted Np protein composed from 590aa. In COS7 cell system, full and regional Gp gene on pDsRed vector induced fluorescence of red in about 30% cells of all, which indicated that express Gp gene. In spites of doing and expressing on the vectors, no amount of the proteins was good enough for diagnosis of LCMV infection in mice. In E.coli (BL21), three peptide fragments of Np was express, but the reactivity with anti-sera was not so high.
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