Estimation of the mutation rates of various genetic DNA markers on Y-chromosome and searching for its recombination hot-spot

Y染色体上各种遗传DNA标记突变率的估计及重组热点的寻找

• 批准号: 13670422
• 负责人: YAMAMOTO Toshimichi
• 金额: $ 2.3万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670422/
• 关键词: Y-chromosome; STR; Mutation; Genetic polymorphism; Genetic markers

In forensic fields, short tandem repeat loci (Y-STRs) and single nucleotide polymorphisms (Y-SNPs) on Y-chromosome have been used for paternal relationship testing and for detecting genotypes originated from males in sexual assault cases. The haplotypes at 14 Y-STR loci were analyzed about two regional Japanese populations (207 in Honshu and 87 in Okinawa) and a Thai population (117) using the 10-plex (DYS436, DYS439, DYS435, DYS19, DYS460, Y-GATA-H4, DYS391, DYS392, DYS438, DYS437) devised by NIST in the USA and the commercially released Y-PLEX6 kit (DYS393, DYS19, DYS389, DYS390, DYS391, DYS385) having two common loci (DYS19 and DYS391) to the10-plex. The allele frequencies at each locus were calculated, and it was found that DYS436 was very uninformative in all populations. Accordingly, in our modified 10-plex, the locus DYS436 was changed for DYS389I, which is one of the minimal database loci in the worldwide Y-STR Database. Using the modified 10-plex and the Y-PLEX 6,14 Y-STRs were easily typed. The heredity of these 14 Y-STRs from father to the son were analyzed by using 161 Japanese father-son DNA samples, and 5 mutations were observed. Sequence analysis revealed that one repeat was gained at DYS389I, DYS439, Y-GATA-H4 and DYS389II, and one repeat was lost at DYS391 in meiosis. The overall mutation rate was calculated as 0.22%/ locus/ meiosis (95% C.I. 0.09 - 0.55%). This value is about the same as those in German and Norwegian populations. Consequently, the estimation of the mutation rates of DNA markers on Y-chromosome was achieved. Further studies are necessary about searching for the recombination hot-spots.

在法医学领域，Y染色体上的短串联重复序列（Y-STR）和单核苷酸多态性（Y-SNPs）已被用于亲子关系检测和性侵犯案件中男性来源的基因型检测。对日本两个地区人群14个Y-STR基因座的单倍型进行了分析（本州207例，冲绳87例）和泰国人群（117例）使用10-plex（DYS436，DYS439，DYS435，DYS19，DYS460，Y-GATA-H4，DYS391，DYS392，DYS438，DYS 437）和市售的Y-PLEX 6试剂盒（DYS 393、DYS 19、DYS 389、DYS 390、DYS 391、DYS 385），其具有两个共同的基因座（DYS 19和DYS 391）。计算了各位点的等位基因频率，发现DYS 436在所有群体中的信息量很小。因此，在我们的改良的10-plex中，基因座DYS 436被改变为DYS 389 I，DYS 389 I是全球Y-STR数据库中的最小数据库基因座之一。用改良的10-plex和Y-PLEX 6对14个Y-STR进行了分型。利用161份日本人父子DNA样本分析了这14个Y-STR从父亲到儿子的遗传，观察到5个突变。序列分析表明，在减数分裂过程中，在DYS 389 Ⅰ、DYS 439、Y-GATA-H4和DYS 389 Ⅱ处增加了一个重复序列，在DYS 391处丢失了一个重复序列。总突变率计算为0.22%/位点/减数分裂（95% C.I. 0.09- 0.55%）。这一数值与德国和挪威人群中的数值大致相同。从而实现了对Y染色体DNA标记突变率的估计。寻找重组热点还需进一步研究。

项目成果

R.Uchihi: "Haplotype analysis with 14 Y-STR loci using 2 multiplex amplification and typing systems in 2 regional populations in Japan"International J. Legal Medicine. 117. 34-38 (2003)

R.Uchihi：“在日本 2 个地区人群中使用 2 个多重扩增和分型系统对 14 个 Y-STR 位点进行单倍型分析”International J. Legal Medicine。

T. Yoshimoto: "A new triplex STR system without irregular alleles by silver staining and its potential application to forensic analysis"J. Forensic Sciences. 46(3). 448-452 (2001)

T. Yoshimoto：“一种新的三链 STR 系统，通过银染色去除不规则等位基因及其在法医分析中的潜在应用”J.

吉本高士: "新しい蛍光標識quadruplex STRタイピングシステムによる日本人及びタイ人のアリール頻度分布の比較"DNA多型. 9. 167-170 (2001)

Takashi Yoshimoto：“使用新的荧光标记四联 STR 分型系统比较日本人和泰国人的芳基频率分布”DNA 多态性。 9. 167-170 (2001)

M. Mizutani: "Analysis of 168 short tandem repeat loci in the Japanese population, using a screening set for human genetic mapping"J. Human Genetics. 46(8). 448-455 (2001)

M. Mizutani：“使用人类遗传图谱筛选集分析日本人群中的 168 个短串联重复基因座”J.

Usuda K: "Haplotype analysis at 10 Y-STR loci using a novel multiplex PCR amplification and typing system in the Japanese and Thai populations."DNA Polymorphisms. 10. 164-167 (2002)

Usuda K：“在日本和泰国人群中使用新型多重 PCR 扩增和分型系统对 10 个 Y-STR 位点进行单倍型分析。”DNA 多态性。