| Project/Area Number |
10670747
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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 Department of Bioregulation Research, Nagoya City University Medical School, Associate Professor, 医学部, 助教授 (70212462)
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| Co-Investigator(Kenkyū-buntansha) |
KATO Taiji Department of Bioregulation Research, Nagoya City University Medical School, Professor and Chairman, 医学部, 教授 (60094364)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1999: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1998: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | Glia Maturation Factor / Gene / Cloning / グリア細胞成長因子 / GMF / アストロサイト |
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| Research Abstract |
(1) Cloning of human GMF-gamma cDNA
We isolated a human GMF-gamma cDNA from human fetal brain cDNA library. One of the clones contained a 573 nucleotide long insert and an open reading frame of 426 bp encoding a deduced protein of 142 amino acids.
(2) Expression of human GMF-beta and GMF-gamma
The cDNA of human GMF-beta and GMF-gamma were inserted to pAED4 expression vector and introduced to E. coli BL21(DE3). The expressed proteins were purified and subjected to antibody production.
(3) Production of anti-human GMF-beta and GMF-gamma antibodies
Recombinant human GMF-beta and GMF-gamma were used as immunogens for the production of polyclonal antibodies. The each anti-serum recognized human GMF-beta and GMF-gamma respectively, but either of them did not react to rat GMF-beta or GMF-gamma.
(4) Cloning of rat GMF-gamma cDNA
We isolated a rat GMF-gamma cDNA from rat pulmonary artery cDNA library. One of the clones contained a 553 nucleotide long insert and an open reading frame of 426 bp encoding a deduced protein of 142 amino acids.
(5) Expression of rat GMF-beta and GMF-gamma
The cDNA of rat GMF-beta and GMF-gamma were inserted to pAED4 expression vector and introduced to E. coli BL21(DE3). The expressed proteins were purified and subjected to antibody production.
(6) Production of anti-rat GMF-beta and GMF-gamma antibodies
Anti-rat GMF-beta and GMF-gamma serum were produced by the same way of human GMF-beta and GMF-gamma. The anti-sera recognized rat GMF-beta and GMF-gamma respectively.
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