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

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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