Diverse mutations in patients with Menkes disease often lead to exon skipping.
Diverse mutations in patients with Menkes disease often lead to exon skipping.
复制标题
DOI：
--
复制
发表时间：
1994-11
影响因子：
9.8
通讯作者：
S. Das;B. Levinson;S. Whitney;Chris D. Vulpe;Seymour Packman;J. Gitschier
中科院分区：
文献类型：
--
作者：
S. Das;B. Levinson;S. Whitney;Chris D. Vulpe;Seymour Packman;J. Gitschier
通过逆转录-PCR（RT-PCR）和化学裂解错配检测，分析了12例不相关的经典Menkes病患者的成纤维细胞培养物中MNK基因的突变。在10名患者中观察到突变，并且在每种情况下都存在不同的突变。所有的突变将被预测对蛋白质表达具有不利影响。在6名患者中观察到导致剪接异常的突变，仅通过RT-PCR检测，包括两个剪接位点的变化，无义突变，错义突变，小重复和小缺失。其余6例患者的化学裂解分析显示存在一个无义突变，两个相邻的5-bp缺失，和一个错义突变。还观察到缬氨酸/亮氨酸多态性。这些发现，结合先前在15%-20%的Menkes患者中观察到的缺失，表明Southern印迹杂交和RT-PCR将识别大多数患者的突变。
Fibroblast cultures from 12 unrelated patients with classical Menkes disease were analyzed for mutations in the MNK gene, by reverse transcription-PCR (RT-PCR) and chemical cleavage mismatch detection. Mutations were observed in 10 patients, and in each case a different mutation was present. All of the mutations would be predicted to have adverse effects on protein expression. Mutations that resulted in splicing abnormalities, detected by RT-PCR alone, were observed in six patients and included two splice-site changes, a nonsense mutation, a missense mutation, a small duplication, and a small deletion. Chemical cleavage analysis of the remaining six patients revealed the presence of one nonsense mutation, two adjacent 5-bp deletions, and one missense mutation. A valine/leucine polymorphism was also observed. These findings, combined with the prior observation of deletions in 15%-20% of Menkes patients, suggest that Southern blot hybridization and RT-PCR will identify mutations in the majority of patients.