| Project/Area Number |
12357003
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Legal medicine
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| Research Institution | Faculty of Medical Sciences, University of Fukui |
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Principal Investigator |
YASUDA Toshihiro University of Fukui, Faculty of Medical Sciences, Professor, 医学部, 教授 (80175645)
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| Co-Investigator(Kenkyū-buntansha) |
UEKI Misuzu University of Fukui, Faculty of Medical Sciences, Instruction Official, 医学部, 教務職員 (00165656)
JIDA Reiko University of Fukui, Faculty of Medical Sciences, Assistant, 医学部, 助手 (40139788)
KISHI Koichiro Gunma University Graduate School of Medicine, Professor, 医学部, 教授 (30169841)
SAWAZAKI Kazumi Fukui Prefectural Police Headquarters, Scientific Investigation Laboratory, Section Head, 科学捜査研究所, 主任研究官
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| Project Period (FY) |
2000 – 2003
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| Keywords | Persona identification / Forensic application / Deoxyribonuclease / Urine / Genetic polymorphism / Molecular biology / Human genetics / Genetic markers |
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| Research Abstract |
1.We have previously found human deoxyribonuclease(DNase) II to be a genetic marker : DNase II levels in human vary depending on whether the individual has the DNA SF2 *H or *L allele.It was clarified that the A-75G transition in the proximal promoter region causes the allelic difference in the promoter activity of the gene, underlying its genetic polymorphism.
2.We devised a simple DNA extraction procedure suitable for STR typing of urine samples using a commercially available DNA/RNA extraction kit. This method alows urine samples to be kept in both a frozen or aqueous state for forensic use.Furthermore, we devised a procedure that combines a simple extraction method, isoelectric focusing and activity-staining for DNase I typing from aged urine stains.DNase I polymorphism could be considered the first biochemical marker found to be well suited for individualization from small aged urine stains.
3.We confirmed the population genetic basis for DNase I polymorphism in a Japanese populatio
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n for forensic application. Our examination of DNase I types revealed a decreasing north-to-south gradient in the DNA SE1 allele.
4.We discovered a novel variable number of tandem repeat of 56 bp unit in intron 4 of human DNase I gene, resulting in elevation of individualization efficiency of DNase.
5.In order to devise the highly sensitive-and specific-immunological typing for DNase I and II polymorphisms, we succeeded in production of each murine monoclonal anti-human DNase I and DNase II with high specificity using newly developed screening procedure for antibodies.Immunoaffinity procedure using these monoclonal antibodies made the purification of human DNases I and II easier, faster and more effective than the conventional procedure. Furthermore, in order to obtain human DNase I as an immunogen, we developed a procedure consisting of transient expression of the enzyme in transfected mammalian cells with DNase I cDNA and purification of the enzyme from the culture medium.
6.Molecular-evolutional aspects of vertebrate DNase I family have been clarified. Less
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