Development and improvement on practical examination using DNA polymorphisms in the field of Forensic Dentistry

本研究的目的是开发和改进在法医案件中使用高度降解的DNA来源检测DNA多态性的可行和可靠的方法。第一年，我们在日本人群中建立了最详细的mtDNA系统发育，并在211个无亲缘关系的个体中确定了33个单倍群，其中包括3个新谱系。在接下来的几年里，我们进一步研究了79个mtDNA编码区多态性，并基于几乎整个mtDNA基因组的信息，将211个个体划分为197个类型，构成106个单倍群谱系。我们进一步增加了80个日本数据库，并在30个马来人个体中发现了9个新的单倍群谱系。对日本263人进行了Y染色体多态性研究。我们利用40种双等位基因标记建立了日本人群中最详细的Y染色体系统发育，并检测了16种Y- str。这些研究使得利用STR单倍型估计二元单倍群成为可能。我们还发现，高变多态位点47z对估计Y染色体双单倍群非常有用。这些数据表明，mtDNA和Y染色体多态性信息不仅可用于个人身份鉴定，而且可用于法医案件的地理估计。我们研究了模板DNA降解对DNA多态性基因分型的影响，结果表明单核苷酸多态性的检测比STR多态性的检测更可靠。我们进一步研究了唾液蛋白基因家族的多态性，并利用重组多态性蛋白进行了与临床牙科相关的基础研究。如上所述，我们在本研究中表明，snp更可靠地分析高度降解的dna。因此，我们主要增加了mtDNA和Y染色体DNA多态性的snp信息。我们相信，我们已经开发了非常有用的DNA分析方法，为实际法医牙科。

The purpose of this study is developing and improving practicable and reliable methods to detect DNA polymorphisms using highly degraded DNA sources in forensic casework. In the first year, we established the most detailed mtDNA phylogeny in the Japanese population and identified 33 haplogroups including 3 new lineages in 211 unrelated individuals. In the following years, we further investigated 79 coding region polymorphisms of mtDNA and classified 211 individuals into 197 types composed of 106 haplogroup lineages based on the information of almost whole mtDNA genome. We further increased 80 Japanese database and also found 9 new haplogroup lineages in 30 Malay individuals. Y chromosomal polymorphisms were investigated in 263 individuals in the Japanese population. We established the most detailed Y chromosomal phylogeny in the Japanese population using 40 kinds of biallelic markers and also examined 16 kinds of Y-STRs. These studied made it possible to estimate binary haplogroups using STR haplotypes. We also found that the hypervariable polymorphic locus 47z is very informative to estimate Y chromosomal binary haplogroups. These data showed that the information of mtDNA and Y chromosomal polymorphisms was useful not only for personal identification but also for geographic estimation in forensic casework. We investigated the influence of template DNA degradation on the genotyping of DNA polymorphisms and showed that the examination of SNPs is more reliable than that of STR polymorphisms. We further investigated polymorphisms on salivary protein gene family and conducted basic research related to clinical dentistry using recombinant polymorphic proteins. As mentioned above, we showed in this study that SNPs is more reliable for analysis on highly degraded DNAs. Thus we increased information of SNPs mainly on mtDNA and Y chromosomal DNA polymorphisms. We believe that we have developed highly useful methods of DNA analysis for practical Forensic Dentistry.