Kojima T, Yamamoto T, Yoshimoto T, Tamaki K, Huang X L, Ohtaki H, Uchihi R, Armour J A, Chen S, Katsumata Y
Department of Legal Medicine, Nagoya University School of Medicine, Nagoya 466-8550, Japan.
Leg Med (Tokyo). 1999 Sep;1(3):119-26. doi: 10.1016/s1344-6223(99)80023-6.
We evaluated the forensic usefulness of D15S233 (wg1d1), a tetrameric short tandem repeat (STR) locus, in the Japanese and Chinese populations. Typing was performed by denaturing polyacrylamide gel electrophoresis followed by silver staining. Nine different alleles were found in 472 Japanese chromosomes and seven in 186 Chinese chromosomes. 102 alleles sequenced were composed of two kinds of repeats (AGGA and GGGA). All alleles differed in size by one tetranucleotide repeat unit, and no insertion or deletion was found. The expected unbiased heterozygosities in Japanese and Chinese were 0.766 and 0.785, respectively. No significant deviations from the Hardy-Weinberg equilibrium were found in either population. We retyped all samples using an alternative pair of flanking primers in order to detect any spurious appearances of homozygotes due to sequence variation at the primer annealing site. One heterozygous sample had unbalanced density bands when the original primer set was used, but equal density bands when our newly designed primer set was used. Sequencing analysis revealed that the sparser allele had one nucleotide substitution near the 5' end of the annealing site of the original primer region. Thus, all apparently homo/heterozygous samples were thought to be truly homo/heterozygous. We also applied the D15S233 locus to paternity testing and forensic identification. Our results suggest that this locus should be a very useful STR locus for forensic practice in Japanese and Chinese.
我们评估了四聚体短串联重复序列（STR）位点D15S233（wg1d1）在日本和中国人群中的法医学应用价值。采用变性聚丙烯酰胺凝胶电泳及银染法进行分型。在472条日本染色体中发现了9个不同的等位基因，在186条中国染色体中发现了7个。测序的102个等位基因由两种重复序列（AGGA和GGGA）组成。所有等位基因的大小相差一个四核苷酸重复单位，未发现插入或缺失。日本人和中国人的预期无偏杂合度分别为0.766和0.785。在这两个人群中均未发现显著偏离哈迪-温伯格平衡的情况。为了检测由于引物退火位点的序列变异导致的纯合子假阳性现象，我们使用另一对侧翼引物对所有样本进行了重新分型。一个杂合样本在使用原始引物组时出现密度带不平衡，但在使用我们新设计的引物组时出现等密度带。测序分析表明，较稀疏的等位基因在原始引物区域退火位点的5'端附近有一个核苷酸替换。因此，所有明显的纯合/杂合样本被认为是真正的纯合/杂合样本。我们还将D15S233位点应用于亲子鉴定和法医鉴定。我们的结果表明，该位点对于日本和中国人群的法医实践应该是一个非常有用的STR位点。
