Zschocke J, Graham C A, McKnight J J, Nevin N C
Department of Medical Genetics, Belfast City Hospital, Northern Ireland.
Acta Paediatr Suppl. 1994 Dec;407:41-2. doi: 10.1111/j.1651-2227.1994.tb13447.x.
We present a simple, fast, non-radioactive method for the analysis of the polymorphic short tandem repeat (STR) system in the human phenylalanine hydroxylase gene. Previously, sizing of the STR marker involved radiolabelling of PCR amplified fragments and resolution on denaturing polyacrylamide gels using M13 sequencing ladder as a standard. However, this method consistently gave sizes 2 bp longer than the known sequence. The fluorescent method presented here employs internal lane standards and enables accurate sizing of the fragments. To avoid confusion, we suggest that the true fragment lengths are used as reference values in the future. The analysis of STR alleles is valuable for population genetic studies and for targeted mutation screening in phenylketonuria (PKU). It can replace RFLP-based haplotype analysis for carrier detection, and we report its use for prenatal diagnosis in a Northern Irish family with PKU. The analysis of 250 Northern Irish chromosomes, including 128 PKU alleles, showed no significant difference between normal and PKU alleles, with fragment lengths of 238 and 242 bp most common in both groups.
我们提出了一种简单、快速、非放射性的方法，用于分析人类苯丙氨酸羟化酶基因中的多态性短串联重复序列（STR）系统。以前，STR标记的大小测定涉及对PCR扩增片段进行放射性标记，并使用M13测序梯作为标准在变性聚丙烯酰胺凝胶上进行分辨率分析。然而，该方法始终给出比已知序列长2 bp的大小。这里介绍的荧光方法采用内部泳道标准，能够准确测定片段大小。为避免混淆，我们建议今后将真实片段长度用作参考值。STR等位基因分析对于群体遗传学研究和苯丙酮尿症（PKU）的靶向突变筛查很有价值。它可以取代基于限制性片段长度多态性（RFLP）的单倍型分析用于携带者检测，并且我们报告了其在一个患有PKU的北爱尔兰家庭中的产前诊断应用。对250条北爱尔兰染色体（包括128个PKU等位基因）的分析表明，正常等位基因和PKU等位基因之间没有显著差异，两组中最常见的片段长度分别为238 bp和242 bp。
