Forensic Studies on a New Genetic Marker, Human DNase I

新遗传标记人类 DNase I 的法医研究

基本信息

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

项目摘要

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。由于鉴定出人类DNase I多态性的新等位基因DNase1 D1*ièD16，因此可以将DNase I-伴形性分类为21种不同的表型。这些发现允许DNase I-colymormorlism在个人识别中的能力进一步提高。 6型同工酶是第一种标记的类型。2。使用新开发的方法来提取和纯化DNase I从老化和极小的唾液污渍（相当于约10μl唾液）来获得良好的DNase I型结果。因此，已经证实，DNase I-colymormorlism为唾液染色的法医表征提供了有价值的信息3。发现胃癌患者与DNase I表型2的高频之间发现了密切的统计关联。但是，良性胃病和对照组的患者组之间的表型分布没有显着差异。因此，DNASE I 2可能有可能识别P…有更多有藏有或发展胃癌风险的人。4。人们发现，人垂体表现出较高的DNase I酶活性及其基因和下丘脑骑马的表达，可快速且瞬时地诱导DNase I活性的显着升高或下降。因此，这些发现强烈表明DNase I扮演了新型的生物学作用。 Hen和Xenopus laevis dnase的生化和分子生物学特征已经进行了。这些酶与其他脊椎动物DNase的比较允许我们包括后者的DNase I位于独立的位置，远非所有其他酶，远离DNase I-家庭中分子进化的所有其他酶。6。我们以前已经发现人脱氧核糖核酸酶（DNase）II是精液中存在的遗传标记。在这项研究中，我们成功地克隆了编码人类DNase II的完整和基因组DNA。此外，通过鱼类分析实现了基因（DNase2）至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个报道。 DNase II在人类中的水平有所不同，具体取决于个人是否具有DNASE2D1*IE D1H或D1*IE D1L等位基因。 DNase II基因表达的瞬时翻译荧光素酶分析使我们包括代理启动子区域中的A-75G转变会导致基因启动子活性的等位基因差异，从而基于其遗传多态性。8。人DNase II由三个非相同亚基组成。 From site-directed mutagenesis study, it was found that a simple attachment of a carbonhydrate moiety, proteolytic cleavage and the inherit 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测试。较少的