| Project/Area Number |
03660274
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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 |
畜産学(含草地学)
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| Research Institution | UTSUNOMIYA UNIVERSITY |
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Principal Investigator |
YOSHIZAWA Midori UTSUNOMIYA UNIVERSITY,AGRICULTURE,ASSOCIATE PROFESSOR, 農学部, 助教授 (60114162)
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| Project Period (FY) |
1991 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1993: ¥200,000 (Direct Cost: ¥200,000)
Fiscal Year 1992: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1991: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | CATTLE / CHROMOSOME / LOCALIZATION / MAPPING / TRANSGENIC RAT / IN SITU HYBRIDIZATION / FLUORESCENCE IN SITU HYBRIDIZATION / 蛍光in situ hybridization / 高精度分染 / ビオチン / DNA / 遺伝子 / 染色体地図 / Gーバンド |
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| Research Abstract |
The present study aimed to establish a method of localization of genes in chromosomes of domestic animals by in situ hybridization.
Preliminarily, ^3H-labeled LCAT (lecithin-cholesterol acyltransferase)DNA,whose locus has been known(No.16q22), was used for in situ hybridization of human chromosomes as a probe. Then, in order to establish a method of in situ hybridization in bovine chromosomes, ^3H-labeled bovine LDL (low-density lipoprotein)receptor DNA was used as a probe. However, the locus was not determined because this DNA might be not suitable for in situ hybridization as the probe.
Fluorescence in site hybridization(FISH) was attempted to localize a gene on human chromosomes using alpha-satellite DNA probe labeled with biotin. After treatment with avidin-fluorescein isothiocyanate and counterstain with propidium iodide, fluorescent signals were detected on chromosomes. They have shown the loci of alpha-satellite DNA.The established FISH method was applied to localization of a transgene on chromosomes of transgenic rats. Metaphase chromosome spreads were obtained from peripheral blood lymphocyte cultures of transgenic rats, which were produced by insertion of mouse whey acidic protein(m-WAP) into pronuclear rat eggs in YS New Techonology Institute. Chromosomes were G-banded by tripsin technique before FISH.The transgene (m-WAP)was localized at q14 site of no.6 chromosome by karyotype analysis.
It is considered that the present study contributes to the gene mapping in domestic animals.
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