Mutation detection and typing of polymorphic loci through double-strand conformation analysis
Mutation detection and typing of polymorphic loci through double-strand conformation analysis
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DOI：
10.1038/ng0298-192
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发表时间：
1998-02-01
期刊：
影响因子：
30.8
通讯作者：
Madrigal, JA
中科院分区：
文献类型：
--
作者：
Argüello, JR;Little, AM;Madrigal, JA
基因内的变异，如核苷酸取代、缺失和插入，可以影响基因产物的功能，并且在某些情况下是有害的。筛选已知的等位基因变异对于确定疾病和基因关联是重要的。靶向特定突变的技术，如限制酶多态性和寡核苷酸探针或PCR引物反应性，可用于检测特定突变，但这些技术通常不能有效地鉴定新突变。基于核苷酸组成不同的DNA片段在通过聚丙烯酰胺凝胶电泳（PAGE;参考文献2，3）分离后表现出不同的迁移率的原理，已经开发了测量DNA构象变化的方法。在这里，我们描述了一种基于构象的突变检测系统，双链构象分析（DSCA），它提供了一种简单的方法来检测遗传变异和类型复杂的多态性位点，我们展示了应用DSCA检测遗传多态性，如单核苷酸差异的DNA片段长度高达979个碱基对。我们目前的应用DSCA检测囊性纤维化基因（CFTR）的4种不同的突变和131个不同的等位基因编码的HLA I类基因。
Variations, such as nucleotide substitutions, deletions and insertions, within genes can affect the function of the gene product and in some cases be deleterious. Screening for known allelic variation is important for determining disease and gene associations'. Techniques which target specific mutations such as restriction enzyme polymorphism and oligonucleotide probe or PCR primer reactivity are useful for the detection of specific mutations, but these techniques are not generally effective for the identification of new mutations. Approaches for measuring changes in DNA conformation have been developed, based on the principle that DNA fragments which differ in nucleotide composition exhibit different mobilities after separation by polyacrylamide gel electrophoresis (PAGE; refs 2,3). Here we describe a conformation-based mutation detection system, double-strand conformation analysis (DSCA), which provides a simple means to detect genetic variants and to type complex polymorphic loci, We demonstrate the application of DSCA to detect genetic polymorphisms such as a single-nucleotide difference within DNA fragments of up to 979 base pairs in length. We present the application of DSCA in detecting four different mutations in the cystic fibrosis gene (CFTR) and 131 different alleles encoded by HLA class I genes.