Kretz K A, Darby J K, Willems P J, O'Brien J S
Department of Neurosciences, University of California, San Diego, La Jolla.
J Mol Neurosci. 1989;1(3):177-80. doi: 10.1007/BF02918904.
Recently, a subset of fucosidosis patients was identified in which the single EcoRI site in the open reading frame of the human cDNA encoding alpha-L-fucosidase was obliterated. We have employed the polymerase chain reaction technique to amplify alpha-L-fucosidase DNA from the five patients known to carry the EcoRI abnormality as well as four patients and two additional fucosidosis patients who do not carry the EcoRI abnormality. Sequence analysis of the amplified DNA has determined that the EcoRI site was destroyed by a C-T transition in the last position of the EcoRI site. This single base change results in the generation of a stop codon 120 base pairs upstream of the normal stop codon. In addition, we have determined that EcoRI cleavage of amplified DNA may be a useful diagnostic tool in the diagnosis of heterozygotes and in prenatal diagnosis of fetuses at risk for this disease.
最近，在岩藻糖苷贮积症患者中发现了一个亚群，其中编码α-L-岩藻糖苷酶的人类cDNA开放阅读框中的单个EcoRI位点被消除。我们利用聚合酶链反应技术，从已知携带EcoRI异常的5名患者以及4名不携带EcoRI异常的患者和另外2名岩藻糖苷贮积症患者中扩增α-L-岩藻糖苷酶DNA。对扩增DNA的序列分析确定，EcoRI位点的最后一个位置发生了C-T转换，导致该位点被破坏。这一单碱基变化导致在正常终止密码子上游120个碱基对处产生一个终止密码子。此外，我们还确定，扩增DNA的EcoRI切割可能是诊断杂合子以及对有患该病风险的胎儿进行产前诊断的一种有用诊断工具。
