Itoh T, Tanaka T, Nagai R, Kikuchi K, Ogawa S, Okada S, Yamagata S, Yano K, Yazaki Y, Nakamura Y
Human Genome Center, Institute of Medical Science, University of Tokyo, Japan.
Hum Genet. 1998 Sep;103(3):290-4. doi: 10.1007/s004390050819.
To elucidate the role of the KVLQT1 gene in the pathogenesis of long QT syndrome (LQTS), we have established a screening system for detecting KVLQT1 mutations by the polymerase chain reaction-single strand conformation polymorphism technique (PCR-SSCP). We first determined exon/intron boundaries and flanking intronic sequences, and found that the KVLQT1 gene consists of 17 coding exons. Subsequently, we synthesized oligonucleotide primers to cover the coding region and the flanking intronic sequences, and searched for mutations in 31 Japanese LQTS families. When genomic DNA from each proband was examined by PCR-SSCP followed by direct DNA sequencing, mutations were detected in five families; two independent families carried the same mutation and three of the four were novel. Each mutation was present in affected relatives of the respective proband. This work will enable us to search more thoroughly for LQTS mutations associated with KVLQT1, and eventually will help us in finding genotype/phenotype relationships.
为阐明KVLQT1基因在长QT综合征（LQTS）发病机制中的作用，我们建立了一种利用聚合酶链反应-单链构象多态性技术（PCR-SSCP）检测KVLQT1突变的筛查系统。我们首先确定了外显子/内含子边界及侧翼内含子序列，发现KVLQT1基因由17个编码外显子组成。随后，我们合成了覆盖编码区及侧翼内含子序列的寡核苷酸引物，并在31个日本LQTS家系中寻找突变。当通过PCR-SSCP随后直接进行DNA测序检测每个先证者的基因组DNA时，在5个家系中检测到了突变；两个独立家系携带相同突变，4个突变中有3个是新发现的。每个突变都存在于各自先证者的患病亲属中。这项工作将使我们能够更全面地寻找与KVLQT1相关的LQTS突变，并最终有助于我们发现基因型/表型关系。
