Forensic Studies on a New Genetic Marker, Human DNase I

新遗传标记人类 DNase I 的法医研究

• 批准号: 10307010
• 负责人: KISHI Koichiro
• 金额: $ 23.87万
• 依托单位: Gunma University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10307010/
• 关键词: 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

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 … More 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

1.由于人类DNase I多态性的新等位基因DNASE1ィイD1*ィエD16被鉴定出来，DNase I多态性可分为21种不同的表型。这些发现使得DNase I多态性在个人身份识别中的能力进一步提高。 6型同工酶是第一个被证明不稳定的同工酶。2．使用新开发的方法从老化的和极小的唾液污渍（相当于约10μl唾液）中提取和纯化DNase I，获得了良好的DNase I分型结果。因此，证实DNase I多态性为唾液染色的法医鉴定提供了有价值的信息。 3．胃癌患者与高频率的 DNase I 表型 2 之间存在密切的统计相关性。然而，良性胃病患者组和对照组之间的表型分布没有显着差异。因此，DNase I 2 可能具有识别患有或发展为胃癌风险的患者的潜力。4。研究发现，人脑下垂体表现出较高的 DNase I 酶活性，其基因和下丘脑激素的表达以快速且短暂的方式诱导 DNase I 活性显着升高或下降。因此，这些发现强烈表明 DNase I 发挥除消化作用之外的新生物学作用。5。已经对母鸡和非洲爪蟾 DNase I 进行了生化和分子生物学表征。通过将这些酶与其他脊椎动物的DNase I进行比较，我们得出这样的结论：从DNase I家族分子进化的角度来看，后者的DNase I处于远离所有其他酶的独立位置。6．我们之前发现人类脱氧核糖核酸酶 (DNase) II 是精液中存在的遗传标记。在这项研究中，我们成功克隆了编码人类 DNase II 的互补 DNA 和基因组 DNA。此外，通过 FISH 分析实现了基因 (DNASE2) 到 19p13.2-p13.1 的区域定位。这是哺乳动物 DNase II.7 的 cDNA 和基因的克隆和表征的第一份报告。人类的 DNase II 水平取决于个体是否具有 DNASE2ィイD1*ィエD1H 或ィイD1*ィエD1L 等位基因。 DNase II 基因表达的瞬时转染荧光素酶分析使我们得出这样的结论：近端启动子区域的A-75G 转换导致该基因启动子活性的等位基因差异，这是其遗传多态性的基础。8．人类 DNase II 由三个不同的亚基组成。通过定点诱变研究，发现酶的亚细胞分选和成熟可能需要同时附着碳水化合物部分、蛋白水解裂解和固有信号肽。考虑到本研究项目中的这些发现，DNase I 已被证实是最有用和最有效的遗传标记之一，适用于法医目的，例如个人身份识别和鉴定。 亲子鉴定。较少的

项目成果

Toshihiro Yasuda: "Molecular cloning of the cDNA encoding human deoxyribonuclease II"Journal of Biological Chemistry. 273(5). 2610-2616 (1998)

Toshihiro Yasuda：“编码人脱氧核糖核酸酶 II 的 cDNA 的分子克隆”生物化学杂志。

安田 年博: "ヒトdeoxyribonuclease II(DNase II)に関する最近の研究"北関東医学. 49(1). 39-41 (1999)

Toshihiro Yasuda：“人类脱氧核糖核酸酶 II (DNase II) 的最新研究”Kitakanto Igaku 49(1) (1999)。

T. Yasuda, H. Takeshita, R. Iida, S. Tsutsumi, T. Nakajima, O. Hosomi, S. Mori, K. Kishi.: "Structure and organization of the human deoxyribonuclease II (DNase II) gene"Ann. Hum. Genet.. 62(4). 299-305 (1998)

T. Yasuda、H. Takeshita、R. Iida、S. Tsutsumi、T. Nakajima、O. Hosomi、S. Mori、K. Kishi.：“人类脱氧核糖核酸酶 II (DNase II) 基因的结构和组织”Ann。

Haruo Takeshita: "Postmortem absorption of dichloromethane: a case study and animal experiments"International Journal of Legal Medicine. (in press)

Haruo Takeshita：“二氯甲烷的死后吸收：案例研究和动物实验”国际法律医学杂志。

Tamiko Nakajima: "Two novel screening methods for selecting monoclonal antibodies which specifically inhibit DNase I enzyme activity" Immunol.Invest.27(3). 145-152 (1998)

Tamiko Nakajima：“用于选择特异性抑制 DNase I 酶活性的单克隆抗体的两种新筛选方法”Immunol.Invest.27(3)。