| Project/Area Number |
12670767
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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 |
Pediatrics
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| Research Institution | Nagoya City University |
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Principal Investigator |
ASAI Kiyofumi Nagoya City University Graduate School of Medical Sciences, Professor, 大学院・医学研究科, 教授 (70212462)
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| Co-Investigator(Kenkyū-buntansha) |
MIURA Yutaka Nagoya City University Graduate School of Medical Sciences, Associate Professor, 大学院・医学研究科, 助教授 (90285198)
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| Project Period (FY) |
2000 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 2002: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2001: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2000: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | Glia Maturation Factor / Enzyme Immunoassay / Genomic DNA / Brain Development / Glia Maturation Factor Beta / Glia Maturation Factor Gamma / グリア成長因子 / EIA / グリア細胞成長因子 |
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| Research Abstract |
(1) Production of Specific Antibodies against Glia Maturation Factor beta (GMFB) and gamma (GMFG).
Recombinant proteins of rat GMFB and GMFG were expressed by E. coli (BL21(DE3)) using pAED4 expression vector, and purified by column chromatography. Polyclonal antibodies were obtained by immunization to New Zealand White Rabbits. Specific antibodies were isolated by affinity chromatography. By western blot, no cross-reactivity was detected between GMFB antibody and GMFG, or between GMFG antibody and GMFB.
(2) Establishment of Enzyme Immunoassay System for GMFB and GMFG.
Sandwich EIA system was established by using the specific antibodies against GMFB and GMFG respectively. These EIAs enabled us to identify immunoreactive GMFB and GMFG in human plasma and rat organs. GMFB was expressed in brain, spleen, thymus, lung and intestine, and GMFG was expressed in thymus and spleen. These results correspond to the results of western blot and immunohistochemical analyses.
(3) Cloning of genomic DNA of human and mouse GMFG.
Genomic DNA of human and mouse GMFG were cloned. Sequence analyses revealed that both human and mouse GMFG gene were consisted by 7 exons.
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