分子克隆技术制备D5S2845基因座的等位基因分型标准物及其法科学应用研究

目的:调查D5S2845基因座在华东汉族人群的群体遗传学数据,解决长期困扰短串联重复序列(shorttandem repeat,STR)分型上存在的准确性和标准化问题。方法:首先利用PCR结合聚丙烯凝胶电泳技术调查华东汉族群体的等位基因频率及基因型分布,再用PCR扩增出D5S2845基因座的8个等位基因片段,将其插入pUC重组质粒中,经DNA测序分析证实插入片段的结构及大小,用国际标准将插入的等位基因片段进行命名,最后经转染、扩大培养、扩增及再鉴定后,制备出标准的D5S2845等位基因分型标准物。结果:D5S2845基因座在华东汉族群体中均有较高的多态性,其非父排除概率及个人识别能力分别为0.558和0.896,应用分子克隆技术制备出大量的D5S2845基因座等位基因分型标准物。结论:D5S2845基因座是一个适合于群体遗传学研究和法科学应用的遗传标记。分子克隆方法制备的STR基因座等位基因分型标准物在法科学实践中应用价值较高。     

华东地区汉族群体20个插入缺失多态的群体遗传学研究及法医学应用

目的:筛查适合华东汉族群体法医学应用的插入缺失多态并进行群体遗传学研究.方法:运用PCR技术结合聚丙烯酰胺凝胶电泳方法对筛选的插入缺失多态进行分型并通过统计分析获得其相关法医遗传学参数.结果:筛选出了20个适合华东汉族群体法医学应用的插入缺失多态位点,并获得了这些基因座的等位基因频率.该20个基因座都具有较好多态性,其中最小等位基因频率为0.108,经计算该20个基因座的累积个人识别机率为0.999 993 845,累积非父排除率为0.916.从20个插入缺失多态位点中我们进一步筛选出了11个最小等位基因频率＞0.3的位点,对这11个具高信息量插入缺失位点进行进一步分析,结果显示11个插入缺失位点累积个人识别机率达到0.996 258 773,累积非父排除率达到0.814 27.结论:本研究获得的上述插入缺失多态位点等位基因频率丰富了华东汉族群体的群体遗传学资料,为插入缺失多态的法医学应用提供了候选的基因座.     

Lack of association between ADRA2B -4825 gene insertion/deletion polymorphism and migraine in Chinese Han population

Objective

The present study aimed to estimate the association between susceptibility to migraine and the 12-nucleotide insertion/deletion (indel) polymorphism in promoter region of α_2B-adrenergic receptor gene (ADRA2B).

Methods

A case-control study was carried out in Chinese Han population, including 368 cases of migraine and 517 controls. Genomic DNA was extracted from blood samples, and DNA fragments containing the site of polymorphism were amplified by PCR. Data were adjusted for sex, age, migraine history and family history, and analyzed using a logistic regression model.

Results

There was no association between indel polymorphism and migraine, at either the allele or the genotype level.

Conclusion

These findings do not support a functional significance of ADRA2B indel polymorphism at position -4825 relative to the start codon in the far upstream region of the promoter in the present migraine subjects.