45例国人Wilson病基因第8外显子突变热区检测和序列分析  

Detection Hot Spots and Sequence-Analyzing the Mutation of Exon Eight in Wilson Disease Gene in 45 Chinese People

作　　者：林豪杰[1] 李乃忠[2] 董强[2] 汪昕[1] 左伋[3] 

机构地区：[1]复旦大学附属中山医院神经内科,上海200032 [2]复旦大学附属华山医院神经内科,上海200040 [3]上海医学院细胞与遗传医学系,上海200032

出　　处：《复旦学报（医学版）》2003年第3期196-199,共4页Fudan University Journal of Medical Sciences

基　　金：卫生部科研基金(961184)

摘　　要：目的 检测中国人群 Wilson病基因(ATP7B)第8外显子突变频率并进行序列分析。方法 采用聚合酶链反应-单链构象多态性(PCR-SSCP)银染技术初步研究了30例患者和15例正常人ATP7B基因第8外显子突变频率,然后对突变个体直接测序,进而与临床表型作相关分析。结果 SSCP发现有6例患者出现第8外显子泳动异常(20％,6/30),且均为杂合子;4例患者既存在2 273G→T,又同时伴有2 250C→G及 2 280G→T;前者是错义突变(即Arg778Leu),累及Tm4区,后者由于氨基酸组成均为Leu,仅是一种保守改变,对Tm4区无影响;1例患者同时存在2099A→G、2 273G→T、2250C→G及2 280G→T4个位点突变,前两者分别影响Tm3、Tm4区,后两者认为是保守改变;1例患者 2 245G insertion是一种新的突变类型,影响Tm4区。结论 提示中国人 ATP7B基因第8外显子可能为一突变热区,2 273G→T(Arg 778Leu)是一种常见的突变类型,Tm4是常见受累功能区之一。Purpose: To detect and analyze the mutation frequency of the exon 8 of ATP7B gene in Chinese patients with Wilson disease. Methods: We preliminarily studied the mutation frequency of the exon 8 of ATP7B gene using a polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and silver staining technique in sample of 30 patients with Wilson disease and 15 normal controls. Then we made correlative analysis between mutations and phenotypes. Results: Abnormal migration bands in PCR-SSCP were observed in 6 heterozygote of 30 patients (20%, 6/30), among them, 4 patients presented missense mutations with 2 273G&rarrT (Arg778Leu), which disrupted Tm4 domain of ATPase7B, and simultaneously with synonymous mutations in 2 250C&rarrG and 2 280G&rarrT. One patient existed four mutation motifs, ie, 2 099A&rarrG(Lys720Arg) with missense mutation and Tm3 domain disrupted;2 273G&rarrT(Arg778Leu) with missense mutation and Tm4 domain disrupted; 2 250C&rarrG and 2 280G&rarrT, with the same amino-acid Leu, were considered conservative changes; Another insertion mutation with 2 245G(769inser His) was also detected, which changed amino acid sequences and disrupted Tm4 domain. Conclusions: Exon 8 might be a hot spot of mutation in ATP7B gene in Chinese. 2 273G&rarrT (Arg778Leu) is one common mutation type. Tm4 is one of the most frequent disrupted domains.

关 键 词：患者 外显子 WILSON病 突变 常见 ATP7B基因 改变 序列分析 直接测序 SSCP 

分 类 号：R742.4[医药卫生—神经病学与精神病学] R596[医药卫生—临床医学]