| Project/Area Number |
09671090
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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 |
Hematology
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| Research Institution | KITASATO UNIVERSITY |
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Principal Investigator |
HIGASHIHARA Masaaki Kitasato Univ. Sch. Med. Professor, 医学部, 教授 (80165084)
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| Co-Investigator(Kenkyū-buntansha) |
AKIYAMA Nobu Kitasato Univ. School of Med. Res. Asso. Prof., 医学部, 助手 (50286358)
KANEKO Takako Kitasato Univ. School of Med. Assis. Prof., 医学部, 講師 (00286361)
KAMATA Kouju Kitasato Univ. School of Med. Assis. Professor, 医学部, 講師 (20146343)
宮崎 浩二 東京大学, 医学部, 医員 (90261966)
須永 真司 東京大学, 医学部, 医員 (70282621)
堀江 良一 東京大学, 医学部, 助手 (80229228)
青木 克已 東京大学, 医学部, 助手 (40291322)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1998: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1997: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | myosin / myosin I / cloning / platelet / cord blood / sequence / cDNA / library / mpl / 網血小板 / 血液細胞 / モノクローナル抗体 |
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| Research Abstract |
We had made the cDNA libraries of human cord blood cells, K562 (erythroleukemia cell line) and Meg-01 (megakaryocytic cell line). Furthermore, we established the method of calculation of "retiplatelet" for the evaluation of megakaryocyte-platelet production, using flow cytometric analysis. To make cloning of novel myosin, we made random primer of myosin I. Finally we had got the 4 clones using K562 library as template. By homology-analysis we found one is myosin VI, another myosin IX, and two unknown clones. RT-PCR method revealed that one of these two clones was also found in CMK and Meg-01, both of which are megakaryocytic cell lines. These results suggest that this clone is a novel unconventional myosin. This novel myosin is constituted in cultured cell from human organs including peripheral blood cells and bone marrow cells.
We are now searching new myosin using the library of human cord blood cells, about 10,000 clones of which were already sequenced.
Another our projects 1) sequencing both 20-kDa and 17-kDa myosin light chains, 2) functional analysis of conventional myosin heavy chain IIA and IIB in blood cells, are also underway.
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