Kordicheva S Iu, Rubtsova G A, Shitova M V, Shaĭkhaev G O, Afanas'ev K I, Zhivotovskiĭ L A
Genetika. 2010 Aug;46(8):1143-7.
Population studies with the use of microsatellite markers face a problem of null alleles, i.e., the absence of a PCR product, caused by the mutations in the microsatellite flanking regions, which serve as the sites of primer hybridization. In this case, the microsatellite primer associated with such mutation is not amplified, leading to false homozygosity in heterozygous individuals. This, in turn, results in biased population genetic estimates, including the excess of homozygotes at microsatellite loci. Analysis of the population structure of a Pacific salmon species, chum salmon (Oncorhynchus keta Walbaum), revealed the presence of null alleles at the Oke3 microsatellite locus in the population samples, in which an excess of homozygotes was observed. The analysis was performed using different combinations of modified primers chosen to match the Oke3 locus. The use of these primers enabled identification of true heterozygotes among those individuals, which were previously diagnosed as homozygotes with the use of standard primers. Removal of null alleles eliminated the excess homozygotes in the chum salmon samples described. In addition to the exclusion of false homozygosity, the use of modified primers makes it possible to introduce polymorphic primer variants associated with certain microsatellite alleles into population studies.
使用微卫星标记的群体研究面临无效等位基因的问题,即由于微卫星侧翼区域(作为引物杂交位点)发生突变而导致PCR产物缺失。在这种情况下,与这种突变相关的微卫星引物无法扩增,导致杂合个体出现假纯合性。这进而导致群体遗传估计出现偏差,包括微卫星位点纯合子过多。对太平洋鲑鱼物种大麻哈鱼(Oncorhynchus keta Walbaum)的群体结构分析表明,在群体样本的Oke3微卫星位点存在无效等位基因,其中观察到纯合子过多。使用选择与Oke3位点匹配的不同组合的修饰引物进行了分析。使用这些引物能够在那些先前使用标准引物诊断为纯合子的个体中识别出真正的杂合子。去除无效等位基因消除了所述大麻哈鱼样本中过多的纯合子。除了排除假纯合性之外,使用修饰引物还使得有可能将与某些微卫星等位基因相关的多态性引物变体引入群体研究。