Forensic molecular biological studies of sex chromosomal DNA for the application to criminal and civil identification

1.通过检测Y染色体特异性α-重复序列进行性别鉴定我们建立了PCR灵敏扩增Y染色体特异性α-重复序列的条件，并成功地从2cm长的毛干中进行了性别鉴定. Y染色体微卫星多态性的检测居住在日本冈山和广岛县的人群中，微卫星DYS 19位点的等位基因频率分别为A=0.064，B=0.068，C=0.423，D=0.226，E=0.211，F=0.004和C-2=0.004（N=263）。在这些等位基因中，C-2等位基因在电泳凝胶上在B和C等位基因之间迁移，尚未在所有种族中报道。通过对家庭样本的检测，证实了STR系统在亲子鉴定中的有效性。此外，这一STR系统有助于查明性攻击的攻击者。Y染色体上的其他STR基因座DYS 390、DYS 391和DYS 395在亲子鉴定和性侵犯案件中也有一定的应用价值. X染色体微卫星HPRT多态性的检测居住在日本冈山县的日本人X染色体上HPRT位点的微卫星等位基因频率估计为T1=0.105，T24=0.243，T3=0.396，T4=0.200和T5=0.057（男性=84，女性=63）。该STR基因座可用于亲子鉴定.釉原蛋白基因的检测我们通过限制性酶分析和测序检查了来自各种动物的PCR产物，包括使用人釉原蛋白序列的引物扩增的人类男性和女性，并且表明除了动物在遗传学上接近之外，这些动物之间的区分是可能的。此外，使用荧光染料标记的引物，可以通过PCR高灵敏度地确定人类男性和女性。

1. Sex determination by detection of Y chromosome-specific aiphoid repeat sequenceWe have established the conditions for sensitive amplification of the Y chromosome-specific aiphoid repeat sequence by PCR, and succeeded sex determination from a hair shaft of 2 cm in length.2. Detection of Y chromosomal microsatellite polymorphismsThe allele frequency of a microsatellite DYS 19 locus was estimated to be A=0.064, B=0.068, C=0.423, D=0.226, E=0.211, F=0.004 and C-2=0.004 (N=263) in Japanese living in Okayama and Hiroshima Prefectures. In these alleles, a C-2 allele, migrated between B and C alleles on electrophoretic gels, have not yet been reported in all the races. By the examination of family samples, the usefulness of this STR system in paternity tests was confirmed. Moreover, this STR system was useful for the identification of an assailant/assailants of a sexual assault. The other STR loci on the Y chromosome, DYS 390, DYS391 and DYS395 were also suggested to be useful in paternity tests and in sexual assault cases.3. Detection of an X chromosome microsatellite HPRT polymorphismThe allele frequency of a microsatellite at HPRT locus on the X chromosome was estimated to be T1=0.105, T24=0.243, T3=0.396, T4=0.200 and T5=0.057 (male=84, female=63) in Japanese living in Okayama Prefecture. This STR locus was suggested to be useful in paternity tests.4. Detection of the amelogenin geneWe have examined PCR products from various animals including human male and female amplified using primers for a human amelogenin sequence by restriction enzyme analysis and sequencing, and it was suggested that discrimination between those animals could be possible except that the animals were close phylogenetically. Moreover, using fluorescent dye-labeled primers, human male and female could be determined with high sensitivity by PCR.