| Project/Area Number |
12680815
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Laboratory animal science
|
|---|
| Research Institution | Nagasaki University |
|---|
Principal Investigator |
OHSAWA Kazutaka Laboratory Animal Center for Biomedical Research, Nagasaki University, Assistant Prof., 医学部, 助手 (90244756)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
KOND Takahito Atomic Bomb Disease Institute, Radiation Effect Research Unit, Biochemistry and Molecular Biology in Desease, Prof., 医学部, 教授 (00158908)
KATNAMINE Shigeru Department of Molecular Microbiology and Immunology, Division of Molecular and Clinical Microbiology, Prof, 大学院・医学研究科, 教授 (40161062)
SATO Hiroshi Laboratory Animal Center for Biomedical Research, Nagasaki University, Prof., 医学部, 教授 (50072947)
|
|---|
| Project Period (FY) |
2000 – 2001
|
| Project Status |
Completed (Fiscal Year 2001)
|
| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2001: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 2000: ¥1,300,000 (Direct Cost: ¥1,300,000)
|
|---|
| Keywords | LCMV / Recombinant antigen / 診断用組換え抗原 / LCMウイルス / 遺伝子組換え抗原 |
|---|
| 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 65.9 kDa, 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. This study left a problem for producing of recombinant antigens with meaningful sensitivity and spescificity.
|
