Racial discrimination by direct sequencing of human mitochondrial DNA

从几个亚洲人群(日本、中国、蒙古、缅甸、台湾、马来西亚和孟加拉国)的200个样本中对人类线粒体DNA高变区进行了测序。从200份唾液中提取DNA，用5%的螯合剂溶液。用引物L15996和H408进行扩增。用染料-引物法进行循环测序反应后，用DNA测序仪(ABI PRISM；型号377)荧光检测高变区的序列变异。我们确定了序列变异的分布和频率，并在最高可变区(16158-16192)的35bp处选择了12个核苷酸。在这些序列中，我们划分为主要的三种类型和三种变种类型。每种类型的频率在亚洲人群中是不同的。在日语中，以Anderson‘s基本型最常见，占65.7%。在马来西亚和缅甸人群中发现了独特的变异。我们正在用Realable方法调查真实的多态位点和多态频率。因此，我们的研究结果显示，在亚洲人群中也发现了几乎相似的多态模式。此外，我们还可以发现，mt-DNA测序并不总是在相同的人群中具有高度的区分性，而是在不同种族来源的人之间提供更多的信息。

Hypervariable region of the human mitochondrial DNA was sequenced from 200 samples in several asian populations (Japan, China, Mongolia, Myanmar, Taiwan, Malaysia, and Bangladesh). DNA were extracted from 200 salivas in solution of 5% chelex. PCR were performed using primers L15996 and H408. The sequence variants of the hypervariable regions were detected by fluorescence with a DNA-sequencer (ABI PRISM ; model 377) after cycle sequencing reaction using dye-primer method. We identified distribution and frequency of sequence variations and selected 12 nucleotide in 35bp of the highest variable regions (16158-16192). In these sequence, we devided major three types and three variants types. Frequency of each types were different among asian populations. In Japanese, Anderson's basic type was the most frequent of 65.7%. Unique variation were encountered in in Malaysian and Myanmar populations.We are investigating of true polymorphic site and frequency of polymorphism with realible method. As a results, our findings show almost similar polymorphic pattern was found within Asian population. Then we can grouped major 5 types of mtDNA.Also, we can find that mt-DNA sequencing is not always highly discriminative among the same populations, but is more informative between people from different racial origins.