| Project/Area Number |
01570340
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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 |
Legal medicine
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| Research Institution | University of the Ryukyus |
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Principal Investigator |
NAGAMORI Hajime University of the Ryukyus, School of Medicine, Department of Legal Medicine, Professor, 医学部, 教授 (50045605)
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| Co-Investigator(Kenkyū-buntansha) |
NAGAMINE Masaru University of the Ryukyus, School of Medicine, Second Department of Biochemistry, 医学部, 助手 (20189161)
KAJIWARA Masahiro Kyorin University, School of Medicine, Department of Legal Medicine, Instructor, 医学部, 助手 (30152656)
UCHIMA Eikoh University of the Ryukyus, School of Medicine, Department of Legal Medicine, Ins, 医学部, 助手 (90093377)
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| Project Period (FY) |
1989 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1990: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1989: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | Individual identification / Paternity testing / DNA polymorphism / Southern hybridization / DNA fingerprinting / Biotinylated probe / DNA指紋法 / サザントランスファ- |
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| Research Abstract |
Jeffreys and his colleagues have developed a DNA fingerprinting system utilizing a variable number of tandem repeats. They produced minisatellite probes which were able to simultaneously hybridize to a number of highly variable DNA regions. The Southern hybridization patterns obtained with these probes consist of multiple hypervariable fragments and are specific to individuals.
DNA fingerprinting has become a powerful new method for forensic applications including individual identification and paternity testing. This method is restrictive in its application to routine work because an isotopically labeled probe is required. Thus we attempted the application of a non-radioactive biotinylated RNA probe derived from SP6 system. However, we have failed to obtain clear fingerprinting due to the lack of sufficient sensitivity. So we have changed another labeling method. The 33mer oligonucleotide corresponding to the repeat unit of the human myoglobin minisatellite was synthesized by the solid phase phosphoramidite method. Biotin was covalently attached via hexylamine linker like an additional base to the 5'end. DNA fragments digested with restriction enzyme were electrophoresed, blotted on nitrocellulose membrane, hybridized with the biotinylated oligonucleotide probe and washed under mild condition. A commercially available kit was used for DNA detection. It was proved that this fingerprinting was comparable to the detection by radioactive method. Therefore, this method may be available for paternity testing and forensic application.
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