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.自从一个新的人类DNA酶I多态性等位基因，DNA酶1等位基因D1* 等位基因D16被发现，DNA酶I多态性可分为21种不同的表型。这些发现允许DNA酶I多态性在个人识别中的能力进一步提高。6型同工酶是第一个被证明是不稳定的类型。用一种新的方法从老年人和极微量唾液斑（相当于约10μl唾液）中提取和纯化DNA酶I，获得了良好的DNA酶I分型结果。因此，DNA酶I多态性为唾液斑的法医学鉴定提供了有价值的信息.胃癌患者与DNA酶I表型2的高频率之间存在密切的统计学关联。然而，良性胃病患者组和对照组之间的表型分布无显著差异。因此，DNase I2可能具有鉴定P ...更多信息 有胃癌风险的患者。发现人垂体显示出较高的DNase I酶活性，并且其基因和下丘脑激素的表达以快速和短暂的方式诱导DNase I活性的显著升高或降低。因此，这些发现强烈表明，DNA酶I发挥新的生物学功能以外的消化之一。已进行母鸡和非洲爪蟾DNA酶I的生化和分子生物学表征。将这些酶与脊椎动物中的其他DNA酶I进行比较，从DNA酶I家族分子进化的角度来看，脊椎动物中的其他DNA酶I处于一个独立的位置。我们以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，我们成功地克隆了互补和基因组DNA编码的人DNase II。此外，通过FISH分析，该基因（DNASE 2）区域定位于19p13.2-p13.1。这是哺乳动物DNase Ⅱ.7的cDNA和基因的克隆和鉴定的首次报道。人的DNA酶II水平取决于个体是否具有DNASE 2等位基因D1* D1 D1 H或D1* D1 D1 L等位基因。DNase II基因表达的瞬时转染荧光素酶分析允许我们得出结论，近端启动子区域中的A-75 G转换导致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。人DNA酶II由三个不同的亚基组成。通过定点突变研究发现，DNase I的亚细胞分选和成熟可能需要同时连接一个糖基、蛋白水解和固有的信号肽，因此，DNase I被证实是一种非常有用和有效的遗传标记，适用于法医学领域，如个人识别和亲子鉴定。少

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)。