Holm L E, Loeschcke V, Bendixen C
Department of Animal Breeding and Genetics, Danish Institute of Agricultural Sciences, Tjele, Denmark.
Mar Biotechnol (NY). 2001 Nov;3(6):555-60. doi: 10.1007/s10126-001-0057-7.
The use of microsatellites for studies of population structure, as markers in genome mapping, and for parentage control has become increasingly popular in recent years. However, the presence of null alleles can lead to confounding results when using microsatellites. In the Omy3DIAS microsatellite, the presence of a null allele was discovered by analysis of family material. The null allele was sequenced after amplification with new primers located farther away from the repeat sequence. The null allele was shown to be caused by a deletion of a 4-bp sequence, which was part of a repetitive sequence within one of the primer recognition sites. As this phenomenon has been seen in other cases of null alleles, this observation leads to the recommendation to avoid repetitive sequences of any kind within primer sequences. Allele-specific amplification of the null allele revealed the presence of a single variant of this allele.
近年来,微卫星在群体结构研究、基因组图谱标记以及亲权鉴定中的应用越来越普遍。然而,使用微卫星时,无效等位基因的存在可能会导致混淆结果。在Omy3DIAS微卫星中,通过对家系材料的分析发现了一个无效等位基因。使用位于距重复序列更远位置的新引物进行扩增后,对该无效等位基因进行了测序。结果表明,该无效等位基因是由一个4碱基序列的缺失引起的,该序列是其中一个引物识别位点内重复序列的一部分。由于在其他无效等位基因案例中也观察到了这种现象,因此建议避免引物序列中出现任何类型的重复序列。对该无效等位基因的等位基因特异性扩增揭示了该等位基因单一变体的存在。