| Project/Area Number |
63571082
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
医学一般
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| Research Institution | The Institute of Physical and Chemical Research |
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Principal Investigator |
IMAI Takashi The Institute of Physical and Chemical Research, Gene Bank, Research Scientist, ジーンバンク室, 研究員 (50183009)
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| Project Period (FY) |
1988 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1989: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1988: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | Human chromosome 21 / YAC / Ordered-clones library / 酵母人工染色体ベクター / コンティグマップ / アガロース顆粒法 / Alu配列 / ヒト、ハムスター雑種細胞 / 長鎖DNA |
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| Research Abstract |
The aim of this project is construction of ordered-clones library for human chromosome 21 using the large insert cloning system, yeast artificial-chromosomes (YAG) system.
1. The first YAG library was constructed with DNA isolated from the human X Chinese hamster hybrid cells which contained chromosome 21 as only human chromosome. However, because it seemed to be difficult to confirm whether the chromosome 21 was kept as normal chromosome in the hybrid cells or not, and because the new rapid-method for screening YAC library has developed, it was not necessary to use the hybrid cells as source of DNA- Therefore, the second library was constructed with human DNA isolated from the cells whose karyotype was normal.
2. The improved method for cloning large YACs was developed because the large YAGs would facilitate the screening for chromosome-specific clones and promote to determine the order of the clones.
3. The human-YAG library contained about 14,000 clones whose average size was 380 kb. It was estimated that the total length of YACs in the library was equivalent to 1.7-fold of the length of the human haploid genome.
4. Nine chromosome 21-specific clones were isolated using PCR based screening method and these restriction maps were constructed.
5. Isolation of the end specific DNA-fragments and screening of the overlapped clones are in progress.
Until now, the set of ordered clones which cover whole chromosome 21 have still not isolated. However, the ordered clones library should be constructed in the near future because the YAC library containing large clones has been built and because the rapid and sensitive screening method has been introduced.
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