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采用分子克隆技术大量制备STR基因座等位基因分型标准物 ,解决长期困扰STR分型的准确性和标准化问题。PCR扩增出D12S375基因座的 7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 ,按重复单位的重复次数对插入的等位基因片段进行命名 ,最后经转染、扩增及再鉴定后制备出标准D12S375基因座等位基因分型标准物。应用此分型标准物 ,调查D12S375基因座在成都汉族 ,甘肃回族及新疆维族群体中的基因型分布频率 ,评估其在法医学实践中的应用价值。
Molecular cloning techniques are used to prepare large numbers of STR loci allelic typing standards to solve the problem of accuracy and standardization of long-term STR typing. Seven alleles of D12S375 locus were amplified by PCR and inserted into the pUC recombinant plasmid. The structure and size of the inserted fragment were confirmed by DNA sequencing analysis, and the inserted allele fragments were named according to the repetition number of repeated units Finally, the standard D12S375 locus allelic typing standard was prepared after transfection, amplification and re-identification. Using this typing standard, we investigated the frequency of genotype distribution of D12S375 locus in Chengdu Han, Gansu Hui and Xinjiang Uygur population, and evaluated its application value in forensic practice.