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

性染色体 DNA 的法医分子生物学研究在刑事和民事鉴定中的应用

• 批准号: 09557039
• 负责人: ISHIZU Hideo
• 金额: $ 7.36万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09557039/
• 关键词: Genetic polymorphism; Sexidentification; Sex chromosome; Microsatellite; Alu sequence; Amelogenin gene; Personal identification; Paternity testing

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.

1．Y染色体特异性重复序列检测性别鉴定建立了PCR灵敏扩增Y染色体特异性重复序列的条件，并成功从2cm长的毛干进行性别鉴定。2． Y染色体微卫星多态性检测微卫星DYS 19位点的等位基因频率估计为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）居住在冈山县和广岛县的日本人。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，尚未在所有种族中都有报道。通过对家庭样本的检查，证实了该STR系统在亲子鉴定中的有用性。此外，该 STR 系统对于识别性侵犯的攻击者很有用。 Y 染色体上的其他 STR 位点 DYS 390、DYS391 和 DYS395 也被认为可用于亲子鉴定和性侵犯案件。 3． X染色体微卫星HPRT多态性检测X染色体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）居住在冈山县的日语。该STR位点被认为可用于亲子鉴定。4．牙釉蛋白基因的检测我们通过限制性内切酶分析和测序，检查了来自各种动物（包括人类雄性和雌性）的PCR产物，这些产物使用人类牙釉蛋白序列的引物扩增，并且表明这些动物之间的区分是可能的，除了动物之间的关系很接近之外。系统发育上。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。