摘要
目的检测肝豆状核变性(Wilsondisease,WD)ATP7B基因启动子区的DNA序列,分析其结构,发现存在的突变,通过报告基因瞬时表达研究突变对启动子功能的影响。方法检测36个WD家系71名成员(其中48例WD患者)及20名正常人的基因组DNA序列并进行分析。对发现的突变,进行荧光素酶报告基因瞬时表达研究。结果(1)在正常对照、患者一级亲属和WD患者的启动子区-190、-78和+260位(转录起始点为+1)均发现存在单个碱基的不同;(2)在48例病人中发现3例存在-183位C→T突变,其中2例为纯合突变,另1例为杂合突变,在正常对照、患者一级亲属中未发现此改变;(3)通过荧光素酶报告基因瞬时表达研究,发现-183位C→T突变不影响ATP7B基因启动子的功能。结论本研究在ATP7B基因启动子区未发现存在致病突变,提示ATP7B基因启动子区存在致病突变在中国人群中是不常见的。
Objective To find out the relationship between mutations and pathogenesis of Wilson＇ s disease （WD） by detecting and analyzing the sequence, checking out the mutations of promoter region of WD gene （A TP7B）. Methods DNA from peripheral blood was obtained from 71 subjects of 36 family （48 WD patients, 23 patients＇ first-degree relatives） and 20 normal people were enrolled into the study. DNA sequence of the promoter region of WD gene was analyzed by PCR amplification and direct genomic sequencing. The reporter gene was used to study the mutation influencing the activity of promoter. Results （l） There were three polymorphisms at positions -190, -78, ＋260 （transcription start site as ＋l） of the promoter region ofWD gene. Normal control, WD patients and patients＇ first-degree relatives all present the polymorphisms; （2） 3 of 48 WD patients presented C→T base substitution mutations at the same position -183.2 were homozygous mutation, while the other was heterozygous. Normal control subjects and patients＇ relatives didn＇t show this kind of mutation. （3） Through detecting the luciferase activity, we didn＇t find that C→T base substitution mutation influenced the activity of promoter. Conclusion We did not find out mutations in the promoter region ofA TP7B gene that relate to the pathogenesis of WD, which suggests that mutations in regulatory elements ofA TP7B are uncommon in Chinese patients.
出处
《中华神经医学杂志》
CAS
CSCD
2005年第8期761-765,共5页
Chinese Journal of Neuromedicine
基金
卫生部与中山大学共建"211工程"重点建设项目基金
广东省科技计划项目(2004B33801006)
深圳市科技计划项目(200304246)
关键词
肝豆状核变性
基因
启动子
突变
Hepatolenticular degeneration
Gene
Promoter
Mutation
