摘要
目的建立聚合酶链反应单链构象多态性（ＲＣＲＳＳＣＰ）和银染技术筛查胰岛素受体基因突变方法。方法选择８９例非胰岛素依赖型糖尿病患者，ＰＣＲ扩增编码胰岛素受体酪氨酸激酶域的外显子（外显子１７～２１），进行ＳＳＣＰ电泳，银染色后，对突变样品进行序列分析。结果发现１１例外显子１７和１例外显子２０的异常电泳条带，外显子１７的突变经顺序分析，为ＣＡＣ１０５８→ＣＡＴ１０５８的杂合和纯合多态性突变。结论ＰＣＲＳＳＣＰ结合银染技术是一种操作简便、经济、灵敏度高、适合于临床大样本基因突变筛查的方法。
Objective To establish polymerase chain reaction single strand conformation polymorphism(PCR SSCP) and silver staining technique to screen mutation of insulin receptor gene. Methods 89 patients with NIDDM were collected.Exon 17 21 of insulin receptor gene were amplified by PCR and screened by SSCP electrophoresis.After silver staining,the mutation samples was sequenced.Resulets Eleven variant sequences in exon 17 and 1 in exon 20 were detected. Direct sequence analysis of exon 17 showed that homozygous and heterozygous nonsense mutations were silent polymorphism at position 1058 (CAC→CAT).Conclusion The PCR SSCP combining with silver staining technique is simple, economic, and sensitive.
关键词
糖尿病
受体
胰岛素
单链构象多态性
银染法
Polymorphism, single stranded conformational Receptors, insulin Mutation Polymerase chain reaction
