提高SCA2编码区CAG三核苷酸重复的PCR扩增效率  

Efficacy Improvement of PCR Amplification of CAG Trinucleotide Repeats in the Coding Sequence of Spinocerebellar Ataxia Type II Gene

作　　者：汤熙翔[1] 夏家辉[1] 

机构地区：[1]湖南医科大学医学遗传学国家重点实验室,中国湖南长沙410078

出　　处：《生命科学研究》2000年第3期217-221,共5页Life Science Research

基　　金：国家自然科学基金!( 3 970 0 0 80 )

摘　　要：以遗传性脊髓小脑共济失调 II型基因 ( spinocerebellar ataxia type II gene SCA2 )编码区内的 CAG三核苷酸重复为研究对象 ( G+ C含量为 69.2 % ) ,比较了热启动 PCR、碱基替代 PCR、添加增效剂 ( 1 %～ 1 2 .5%二甲亚砜、1 %～ 2 5%甘油、1 %～ 1 2 .5%甲酰胺 )与常规PCR的扩增效率 ,发现热启动 PCR、碱基替代 PCR及添加增效剂 ( 1 %～ 1 0 %二甲亚砜、5%～ 2 0 %甘油、1 %～ 1 0 %甲酰胺 )能提高该 GC富集区的扩增效率 ,并对 70个 SCA家系及 60个散发 SCA患者进行了 SCA2的基因诊断 .To improve the efficacy of PCR amplification of CAG trinucleotide repeats in the coding sequence of spinocerebellar ataxia type II gene(69.2% G+C), hot start PCR, base replacement PCR, and the addition of enhancers(1%～12.5% DMSO , 1%～25% glycerol ,1%～12.5% formamide) were performed and compared with normal PCR . The results showed that hot start PCR, base replacement PCR and the addition of enhancers(1%～10% DMSO , 5%～20% glycerol , 1%～10% formamide) improved the amplification efficacy of the GC rich region. Gene diagnosis in 70 SCA pedgrees and 60 spontaneous SCA patients were also conducted.

关 键 词：聚合酶链式反应 遗传性脊髓小脑共济失调Ⅱ型 

分 类 号：R744.6[医药卫生—神经病学与精神病学] Q75[医药卫生—临床医学]