Ohshima T, Sasaki M, Takahashi J, Sakuragawa N
Department of Inherited Metabolic Disease, National Institute of Neuroscience, NCNP, Tokyo, Japan.
J Child Neurol. 1994 Jan;9(1):38-40. doi: 10.1177/088307389400900108.
The most common mutation in late-onset metachromatic leukodystrophy is a cytosine-to-thymine substitution in exon VIII. This mutation caused a substitution of leucine for proline at amino acid residue 426. We developed a rapid and simple method for the detection of 426Pro-->Leu mutation by polymerase chain reaction with mismatched primer. Although the 426Pro-->Leu mutation does not alter recognition sequence for restriction enzymes, we created a Pst I restriction site using a 3'-primer mismatched at one nucleotide. As a result, the mutation can be detected as a Pst I restriction fragment length polymorphism.
迟发性异染性脑白质营养不良最常见的突变是外显子VIII中胞嘧啶到胸腺嘧啶的替换。该突变导致第426位氨基酸残基处脯氨酸被亮氨酸取代。我们开发了一种快速简便的方法，通过错配引物聚合酶链反应检测426Pro→Leu突变。虽然426Pro→Leu突变不会改变限制性内切酶的识别序列，但我们使用在一个核苷酸处错配的3'引物创建了一个Pst I限制性位点。结果，该突变可作为Pst I限制性片段长度多态性被检测到。
