| Project/Area Number |
10307010
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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 | 一般 |
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| Research Field |
Legal medicine
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| Research Institution | Gunma University |
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Principal Investigator |
KISHI Koichiro Gunma University School of Medicine, Department of Legal Medicine, Professor, 医学部, 教授 (30169841)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAJIMA Tamiko Gunma University School of Medicine, Department of Legal Medicine, Assistant, 医学部, 助手 (40008561)
TAKESHITA Haruo Gunma University School of Medicine, Department of Legal Medicine, Lecturer, 医学部, 講師 (90292599)
YASUDA Toshihiro Gunma University School of Medicine, Department of Legal Medicine, Associate Professor, 医学部, 助教授 (80175645)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥37,300,000 (Direct Cost: ¥37,300,000)
Fiscal Year 1999: ¥13,900,000 (Direct Cost: ¥13,900,000)
Fiscal Year 1998: ¥23,400,000 (Direct Cost: ¥23,400,000)
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| Keywords | Genetic marker / Deoxyribonuclease I / Deoxyribonuclease II / Personal identification / Genetic polymorphism / Molecular biology / Human genetics / Paternity testing / deoxyribonucleaseI / deoxyribonucleaseII / 分子進化 / 遺伝子 / 脳下垂体 |
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| Research Abstract |
1. Since a new alleles, DNASE1 ィイD1*ィエD16, of human DNase I polymorphism was identified, DNase I-polymorphism can be classified into 21 different phenotypes. These findings permit the ability of DNase I-polymorphism in personal identification to be further elevated. The type 6 isoenzyme was the first type to be shown to be labile.
2. Good DNase I-typing results were obtained using a newly developed method for extraction and purification of DNase I fro aged and extremely small saliva stains (equivalent to about 10μl saliva). Therefore, it was confirmed that DNase I-polymorphism provides valuable information for forensic characterization of saliva stains.
3. A close statistical association was found between patient with gastric carcinoma and a high frequency of DNase I phenotype 2. However, there was no significant difference in the phenotype distribution between the group of patients with benign gastric diseases and controls. Therefore, DNase I 2 may have potential for identification of p
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atients who are at risk of harboring or developing gastric carcinoma.
4. It was discovered that the human pituitary gland exhibits higher DNase I enzyme activity and expression of its gene and hypothalamic hormones induced a significant elevation or decline of DNase I activity in a rapid and transient manner. Therefore, these findings strongly suggest that DNase I plays novel biological roles other than a digestive one.
5. Biochemical and molecular-biological characterizations of hen and Xenopus laevis DNase I have been performed. Comparison of these enzyme with other vertebrate DNase I allowed us to conclude that the latter DNase I is situated at an independent position far from all the other enzymes from the standing point of molecular evolution in DNase I-family.
6. We have previously found human deoxyribonuclease (DNase) II to be a genetic marker present in semen. In this study, we succeeded in cloning of both the complementary and genomic DNAs encoding human DNase II. Furthermore, regional localization of the gene (DNASE2) to 19p13.2-p13.1 was achieved by FISH analysis. This is the first report of the cloning and characterization of the cDNA and gene for mammalian DNase II.
7. DNase II levels in human vary depending on whether the individual has the DNASE2ィイD1*ィエD1H or ィイD1*ィエD1L allele. The transient transfection luciferase analysis of the DNase II gene expression permit us to conclude 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.
8. Human DNase II was composed of three non-identical subunits. From site-directed mutagenesis study, it was found that a simultaneous attachment of a carbohydrate moiety, proteolytic cleavage and the inherent signal peptide might be required for subcellular sorting and maturation of the enzyme.
Considering these findings made in this research project, DNase I has been confirmed to be one of the most useful and effective genetic markers suited for forensic purpose such as personal identification and paternity testing. Less
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