摘要
目的鉴定并分析1个新的肾上腺脑白质营养不良家系的基因突变类型和位点。方法采用长链RT-PCR技术,从外周血提取总RNA并合成cDNA,以cDNA为模板,分4个片段扩增致病基因编码区,将纯化后的PCR产物直接测序,找出基因突变的位点。同时应用限制性酶切分析的方法,分析发病家系所有成员的基因组DNA,以证实所发现的突变。结果患者肾上腺脑白质营养不良基因外显子2上的第343位密码子发生了GGC→GTC改变,使原来编码的甘氨酸被缬氨酸(G343V)取代;患者母亲为G343V突变携带者,患者哥哥的基因型和患者完全相同,其父亲为正常基因型。结论在中国人肾上腺脑白质营养不良患者中发现一个新的ABCD1基因突变,即G343V突变。
Objective To identify the ABCD1 gene mutation in a Chinese family with X-linked adrenoleukodystrophy (ALD). Methods Total RNA was isolated from the peripheral blood of the patient and his mother. After reverse transcription, the coding region of ABCD1 gene cDNA was amplified in 4 segments by PCR. The PCR products were purified and directly sequenced. The result of sequencing was confirmed by restrictive enzyme digestion of PCR products from genomic DNA of patient's family members. Results A missense mutation of GGC→GTC was detected at codon 343 of patient's ABCD1 gene, resulting in the replacement of glycine by valine. The same genotype was found in his elder brother and the heterozygote for this mutation was found as well in patient's mother. Conclusion A novel missense mutation, G343V, was identified in the ABCD1 gene of a Chinese patient with ALD.
出处
《中华神经医学杂志》
CAS
CSCD
2005年第3期241-243,共3页
Chinese Journal of Neuromedicine
