Analysis of hypervariable minisatellites and their flanking sequences towards application to forensics polymorphic markers

一些小卫星位点显示出非常高水平的等位基因长度变异性。大多数小卫星基因座由两种或两种以上细微不同重复单元类型的异质阵列组成（小卫星变异重复序列）。通过MVR映射可以确定散布模式。我们分析了日本人群中高变小卫星CEB1 （D2S90）的特性。长度杂合度为85.0%，等位基因平均长度为69个重复。在小卫星的侧翼发现一个新的单核苷酸多态性（SNP）。等位基因内部MVR结构存在差异，重复序列中存在新的碱基替换/缺失，表明CEB1也是日本人微卫星高变异位点之一。微卫星MS32 （D1S8）也是MVR-PCR显示出巨大多样性的微卫星之一。在调查的几个民族人群中，几乎所有的等位基因都是不同的。然而，不同的等位基因可以在重复组织中表现出显著的相似性。然而，到目前为止，现有的亚洲数据库几乎完全由日本的等位基因组成。为了探索日本人和其他亚洲人等位基因之间的相似性，我们开始了对泰国人DNA样本的MS32等位基因定位。利用等位基因特异性引物对SNP杂合个体进行MVR-PCR，该SNP的等位基因特异性MVR-PCR可定位120个等位基因。在106个泰国等位基因中，只有2对等位基因是不可区分的，其余102个等位基因是唯一的。在无限等位基因模型下，假设选择性中立，这些数据给出了估计的θ。泰国人的价值是2710美元。对多样性的估计也表明杂合度为99.96%。采用启发式点阵分析对等位基因编码进行比较。利用MS32等位基因编码数据库，共包含1072个不同民族人群的等位基因。在泰国的等位基因中，有41个等位基因具有显著的等位基因间相似性，占所有等位基因的39%。与世界等位基因相比，鉴定出57个相似度是真实的，占所有等位基因图谱的54%。在相关的泰国等位基因中，64%的等位基因与日本等位基因有显著的相似性，26%的等位基因与高加索等位基因有显著的相似性。塔希等位基因与非洲等位基因无亲缘关系。因此，泰国人的等位基因在很大程度上是亚洲人特有的，尽管这种趋势似乎不像日本人那样高。MS32的MVR等位基因分析不仅可以作为个体鉴定的工具，还可以作为种族背景的工具，为探索人类种群历史上最近发生的事件提供有用的谱系标记。少

Some minisatellite loci show very high levels of allele length variability. Most minisatellite loci consist of heterogeneous arrays of two or more subtly different repeat unit types (minisatellite variant repeats). Interspersion patterns can be determined by MVR mapping. We analysed the properties of a hypervariable minisatellite CEB1 (D2S90) in the Japanese population. The length heterozygosity is observed at 85.0% and the average length of the alleles is 69 repeats. A new single nucleotide polymorphism (SNP) was found in the flanking the minisatellite. Internal MVR structure was different between alleles and new base substitutions / deletions were found within repeats, which indicates CEB1 is also one of the most hypervariable minisatellite loci in Japanese. Minisatellite MS32 (D1S8) is also one of the minisatellites that shows vast diversity by MVR-PCR. Almost all alleles in several ethnic populations surveyed were found to be different. However, distinct alleles can show significan … More t similarities in repeat organization. However, up until now the existing Asian database has consisted almost entirely of Japanese alleles. In order to explore the similarities between Japanese and other Asian alleles, we have started MS32 allele mapping of DNA samples from Thais. By using allele-specific primers in MVR-PCR of individuals heterozygous for a SNP, 120 alleles can be mapped by the allele-specific MVR-PCR of the SNP. In 106 mapped Thai alleles, there are only 2 pairs of alleles are indistinguishable, and the remaining 102 alleles are unique. Under the infinite allele model and assuming selective neutrality, these data give an estimated theta. value of 2710 for the Thai. This estimate of diversity also suggests a heterozygosity of 99.96%. Allele codes were compared with each other by heuristic dot matrix analysis. MS32 Allele code database consisting 1072 alleles of various ethnic populatioins was also used. Within Thai aleles, 41 showed significant inter-allelic similarities, corresponding to 39% of all mapped alleles. Compared with world alleles, 57 were confirmed their similarities are authentic corresponding to 54% of all mapped alleles. Among the related Thai alleles, 64% of alleles show significant similarities to Japanese alleles and 26% to Caucasian. No Tahi alleles are related to African alleles. Therefore, Thai alleles are largely Asian population-specific, although the tendency does not seem to be as high as in Japanese. MVR allele analysis at MS32 can act as a tool not only for individual identification but also for ethnic background providing useful lineage markers for exploring recent events in human population history. Less