| Project/Area Number |
04044028
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Institution | Tohoku University |
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Principal Investigator |
MIZUNO Shigeki Faculty of Agriculture, Tohoku University, 農学部, 教授 (90112903)
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| Co-Investigator(Kenkyū-buntansha) |
HARATA Masahiko Faculty of Agriculture, Tohoku University, 農学部, 助手 (70218642)
HUTCHISON Nancy Fred Hutchinson Cancer Research Center (U.S.A.), スタッフサイエンテイ
NANCY Hutchi フレッドハッチソンがん研究センター (アメリカ), スタッフサイエンテイ
NANCY J Hutc フレッドハッチンソンがん研究センター, (アメリカ、シアトル), スタッフサイエンティ
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| Project Period (FY) |
1992 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥7,500,000 (Direct Cost: ¥7,500,000)
Fiscal Year 1994: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1993: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1992: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | Chicken / Sex Chromosomes / Z Chromosome / W Chromosome / Gene Mapping / Heterochromatin / Lampbrush Chromosome / Fluorescence In Situ Hybridization (FISH) / 減数分裂期染色体 / ZW染色体 / 卵母細胞 / 減数分裂 / 2価染色体 / ゲノムマッピング |
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| Research Abstract |
The aim of this joint research was to isolate clones from the chicken W and Z chromosome-specific genomic libraries, determine their nucleotide sequences, and to examine their localizations on the lampbrush Z-W bivalent chromosomes by fluorescence in situ hybridization (FISH). The major outcome is as follows.
1) Z chromosome : Lambda gt10 libraries were prepared from a terminal region of the chicken Z chromosome by utilizing argon ion laser micro-irradiation to burn down all other chromosomes, micro-extraction of DNA from the remaining chromosomal region and random PCR amplification using a single unique primer. One hundred clones were selected randomly, mixed and used as probes in the FISH to mitotic chromosomes. Those probes hybridized to the heterochromatic end region of the chicken Z chromosome. A lambda GEM12 genomic library of the female chicken was screened with those mixed probes and a clone, pCHZTH8 was obtained. This clone hybridized to the short loops region and the terminal bow-like loop (TBL) at the heterochromatic end of the Z chromosome. A part of the pCHZTH8 sequence is conserved among the order Galliformes but most of them is unique to the genus Gallus.
2) W chromosome : The same technique as above was applied to obtain W chromosome-specific genomic libraries. Two regions on the chicken W chromosome, CW01 and CW50 each consisting of about 25-kb non-repetitive sequences, were cloned. The CW50 region contains a putative CpG island and the CW01 region contains a sequnce that is conserved widely as W-linked among many avian species, and thus is a useful general probe for sexing avian species.
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