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

尽管个体化在法律医学中是一个非常重要的研究领域，但适合于估计年龄的一些与年龄相关的标志物极其有限。在本研究中，我们检测了衰老和发育的每个阶段的特异性生化标志物，并试图建立通过对这些标志物的生化和分子生物学检测来从法医样本中估计年龄的方法。UGL-Y是一种糖蛋白，被发现是一种年龄相关的标记物，仅在年轻受试者的尿中检测到。在本研究中，我们确定了它的一级结构。UGL-Y由180个氨基酸残基组成，通过数据库检索，UGL-Y的氨基酸序列与部分纤维连接蛋白的氨基酸序列完全一致。根据这一结果，推测UGL-Y可能来源于纤维连接蛋白初级基因转录本的有限酶切产物或选择性剪接形式。我们研究了小鼠k-…中与年龄相关的基因表达。更多的idney来寻找新的与年龄相关的标记。因此，利用荧光差异显示-聚合酶链式反应(FDD-PCR)技术和比较RT-PCR技术可以鉴定出7种与年龄相关的表达基因。因此，这些方法对于检测新的年龄相关差异表达基因是非常有用的。我们已经阐明了上述与年龄相关的基因之一的分子和生物学特性。该基因全长3,058bp，由194个氨基酸残基组成，编码2.2kb蛋白。BLAST搜索表明，该蛋白与过氧化体蛋白MPV 17和PMP 22具有较高的同源性。因此，我们将该新蛋白命名为MPV 17结构域家族的M-LP。此外，该基因全长5kb，有3个外显子，证实了该基因的年龄相关性表达。根据本研究项目的这些发现，我们可以为年龄相关标记的分析和法医应用所必需的分子和生物学方法奠定基础。较少

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