Molecular-biological analysis of age-related markers present in body fluids and its forensic applications

体液中年龄相关标记物的分子生物学分析及其法医学应用

• 批准号: 09357004
• 负责人: KISHI Koichiro
• 金额: $ 18.11万
• 依托单位: Gunma University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A).
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09357004/
• 关键词: estimation of age; age-related markers; gene expression; cDNA cloning; Ugl-Y; Molecular-biological analysis; forensic application; body fluid; PCR; differential display; 分子生物学; 年齢; cDNA; 尿; アミノ酸配列; 鑑識科学

Although individualization in legal medicine is one of very important research field, a number of age-related markers suitable for estimation of age is extremely limited. In this study, we have examined biochemical markers specific to each step of aging and develpment and tried to establish the method to estimate the age from forensic samples by biochemical and molecular-biological detection of these markers1. Ugl-Y is a glycoprotein which has been discovered as an age-related marker detected in urine of only young subjects. In this study, we has determined its primary structure. Ugl-Y consisted of 180 amino acid residues, and from the database search, the amino acid sequence of Ugl-Y was found to be entirely identical to that of a part of fibronectin. From the result, it was assumed that Ugl-Y might be derived from a limited digested product or alternative spliced form of primary gene transcript of fibronectin.2. We have surveyed gene expressed in the age-related manner in the mouse k … More idney to find novel age-related markers. Consequently, seven kinds of age-dependent expressed cDNA could be identified using a fluorescence differential display-PCR (FDD-PCR) technique and comparative RT-PCR technique. Therefore, these methods were found to be very useful for detecting new age-related genes expressed differentially.3. We have elucidated molecular and biological properties of one of age-related genes described above. Its cDNA was 3,058 bp long and consisted of 194 amino acid residues encoding the 2.2 kb protein. From BLAST searching, it was clarified that this protein had high homology to peroxisomal protein, Mpv 17 and PMP 22. Therefore, we named this novel protein as M-LP belonging to Mpv 17 domain family. Moreover, its gene was 5 kb long with three exons, and age-related expression of the gene was confirmed. From these findings made in this research project, we could establish the basis for the molecular and biological approach which is indispensable for analysis and forensic application of age-related markers. Less

尽管法律医学中的个性化是非常重要的研究领域之一，但许多适合年龄的标记估计非常有限。在这项研究中，我们检查了针对每个衰老和发育步骤的生化标记物，并试图通过生化和分子生物学检测这些标记来确定从法医样品估算年龄的方法1。 UGL-Y是一种糖蛋白，已被发现是仅在年轻受试者的尿液中检测到的与年龄相关的标记。在这项研究中，我们确定了其主要结构。 UGL-Y由180个氨基酸残基组成，从数据库搜索中，发现UGL-Y的氨基酸序列与纤连蛋白的一部分完全相同。从结果中，假定UGL-Y可以源自有限的消化产物或纤连蛋白的原代基因转录物的剪接形式。2。我们已经调查了小鼠K中与年龄相关的方式表达的基因……更多的Idney找到了与年龄相关的新标记。因此，可以使用荧光差异显示PCR（FDD-PCR）技术和比较RT-PCR技术来鉴定七种依赖年龄的表达cDNA。因此，发现这些方法对于检测不同年龄相关的基因表达不同非常有用3。我们已经阐明了上述年龄相关基因之一的分子和生物学特性。它的cDNA长3,058 bp，由194个氨基酸组成，编码2.2 kb蛋白。从爆炸搜索中，可以澄清说，该蛋白与过氧化物体蛋白，MPV 17和PMP 22具有很高的同源性。因此，我们将这种新型蛋白命名为属于MPV 17域家族的M-LP。此外，它的基因长5 kb，带有三个外显子，并证实了该基因与年龄有关的表达。从该研究项目中提出的这些发现，我们可以为分子和生物学方法建立基础，这对于与年龄相关标记的分析和法医应用是必不可少的。较少的

项目成果

Haruo Takeshita: "Mammalian deoxyribonuclease I are classified into three types"Biochemical and Biophysical Research Communications. 269(2). 481-484 (2000)

Haruo Takeshita：“哺乳动物脱氧核糖核酸酶 I 分为三种类型”《生物化学和生物物理研究通讯》。

Koichiro Kishi: "DNase I : structure, function, and use in medicine and forensic science"Legal Medicine. (in press). (2001)

岸晃一郎：《DNase I：结构、功能以及在医学和法医学中的应用》法律医学。

Osamu Hosomi: "Molecular cloning of cDNA encoding Xenopus Laevis deoxyribonuclease I"DNA Sequence. (in press).

Osamu Hosomi：“编码非洲爪蟾脱氧核糖核酸酶 I 的 cDNA 的分子克隆”DNA 序列。

Tamiko Nakajima: "Rapid purification of human Dnase I using mouse monoclonal anti-Dnase I antibodies and characterzation of the antibodies"Experimental and Clinical Immunogenetics. (in press).

Tamiko Nakajima：“使用小鼠单克隆抗 Dnase I 抗体快速纯化人 Dnase I 并表征抗体”实验和临床免疫遗传学。

Yasuda,T.: "Chromosomal localization of a human deoxyribonuclease II gene (DNASE2) to 19p13.2-p13.1 using both the polymerase chain reaction and fluorescence in situ hybridization analysis" Biochemical and Biophysical Research communications.

Yasuda,T.：“使用聚合酶链式反应和荧光原位杂交分析将人脱氧核糖核酸酶 II 基因 (DNASE2) 染色体定位到 19p13.2-p13.1”《生物化学和生物物理研究通讯》。