FOR NOVEL GENE-MUTATIONS IN 5 JAPANESE MALE-PATIENTS WITH NEONATAL OR LATE-ONSET OTC DEFICIENCY - APPLICATION OF PCR-SINGLE-STRAND CONFORMATION POLYMORPHISMS FOR ALL EXONS AND ADJACENT INTRONS
FOR NOVEL GENE-MUTATIONS IN 5 JAPANESE MALE-PATIENTS WITH NEONATAL OR LATE-ONSET OTC DEFICIENCY - APPLICATION OF PCR-SINGLE-STRAND CONFORMATION POLYMORPHISMS FOR ALL EXONS AND ADJACENT INTRONS
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DOI：
10.1007/bf00216144
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发表时间：
1993-08-01
期刊：
影响因子：
5.3
通讯作者：
MATSUDA, I
中科院分区：
文献类型：
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作者：
MATSUURA, T;HOSHIDE, R;MATSUDA, I
鸟氨酸转氨基甲酰基酶缺乏症（OTC）是尿素循环中最常见的先天性错误，具有x连锁遗传和频繁的新突变。Southern blots只显示了突变的一小部分，但是利用聚合酶链反应（PCR）扩增cDNA或基因组DNA，然后进行DNA测序，极大地克服了这一困难。剩下的问题是，在前一种情况下，制备完整mRNA的新鲜肝脏样本的可用性有限，而在后一种情况下，只有一些外显子的PCR引物序列。在这里，我们报告了内含子序列的结构，这些序列足够长，可以使用PCR和PCR单链构象多态性（PCR- sscp）分析OTC基因的所有外显子和邻近内含子。我们对5名日本男性新生儿或晚发型患者的发现进行了DNA分析。5名患者在蛋白质编码区有突变。C到G (S192R)， A到T (D196V)， A到G (T264A)， T到C (M268T)，以及C到T （R277W）的替换。其中前四个是新的错义突变，并且在相应的家族中证实了突变的存在。
Ornithine transcarbamylase deficiency (OTC), the most common inborn error of the urea cycle, shows an X-linked inheritance with frequent new mutations. Southern blots reveal only a small percent of the mutation, but amplification of cDNA or genomic DNA using the polymerase chain reaction (PCR) followed by DNA sequencing, has contributed greatly to overcoming this difficulty. Problems remaining are the limited availability of fresh liver samples for preparation of intact mRNA in the former case, and there are primer sequences for PCR for only some exons in the latter case. Here, we report the structures of intron sequences which are long enough to analyze all exons and adjacent introns of the OTC gene using PCR and PCR single-strand conformation polymorphisms (PCR-SSCP). We carried out a DNA analysis of findings in five Japanese male patients with neonatal or late onset form. Five patients had mutations in the protein coding region. C to G (S192R), A to T (D196V), A to G (T264A), T to C (M268T), and C to T (R277W) substitutions. The first four of these were novel missense mutations and the presence of the mutation was confirmed in the corresponding families.