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研究了醛糖还原酶基因转录起始点上游 - 2 1kb处的微卫星DNA标记——— (AC)n二核苷酸串联重复序列 7种等位基因 (Z + 6,Z + 4,Z + 2 ,Z ,Z 2 ,Z 4和Z 6)的筛选方法。首先自人外周血提取基因组DNA后设计一对引物进行PCR扩增 ,获得长度为 13 2～ 14 4bp的DNA片段。选取等位基因为纯合子的个体DNA标本的PCR扩增产物 ,经纯化后直接测序以确定等位基因的种类 ;并以确定的等位基因为标准 ,用尿素 /甲酰胺聚丙烯酰胺凝胶电泳银染显色法对各种等位基因进行筛选。该法操作简便、快速、准确度高 ,利于大量标本的筛选 ,也适用于其它二核苷酸重复序列等位基因的筛选。
In this study, seven alleles (Z + 6, Z + 4, Z +) of microsatellite DNA markers- (AC) n dinucleotide tandem repeat sequence upstream of transcription start point of aldose reductase gene 2, Z, Z 2, Z 4 and Z 6). First, a pair of primers was designed for PCR amplification after extracting genomic DNA from human peripheral blood to obtain a DNA fragment with a length of 13 2 to 14 4 bp. The PCR products of individual homozygous alleles were selected and sequenced directly to determine the type of alleles. Using the defined alleles as the standard, polyacrylamide gel Electrophoresis silver staining for various alleles screening. The method is simple, rapid and accurate, which is conducive to the screening of a large number of specimens. It is also suitable for the screening of other dinucleotide repeat alleles.