G209A突变型α-synuclein基因真核表达质粒的构建  

Construction of an expression system of G209A mutant human α-synuclein

作　　者：李琳[1] 刘振国[1] 陈生弟[1] 倪培华[2] 

机构地区：[1]上海第二医科大学瑞金医院神经内科,上海200025 [2]上海第二医科大学瑞金医院检验科

出　　处：《上海第二医科大学学报》2004年第9期710-712,共3页Acta Universitatis Medicinalis Secondae Shanghai

基　　金：上海市科委青年科技"启明星后"计划(01QMH1410);上海市教委发展基金(03BK24)资助项目.

摘　　要：目的 构建人G209A突变型α-synuclein基因真核表达质粒。方法 采用RT-PCR从人胚脑组织中扩增人α-synuclein基因,再采用SOE法定点突变,构建G209A突变型α-synuclein基因真核表达质粒。结果 酶切和DNA测序证实,突变型(G209A)α-synuclein基因除第209位碱基G被A替代以外,其余序列与野生型完全一致。结论 成功构建G209A突变型α-synuclein基因真核表达质粒,为研究帕金森病发病机制奠定基础。Objective To construct the expression system of human mutant G290A a-synuclein. Methods The cDNA encoding the human α-synuclein was isolated by using RT-PCR method with total RNA extracted from the fetal brain. Mutant G290A α-synuclein was obtained and cloned by gene splicing and overlap extension (SOE) on site-direct mutagenesis of wild-type human α-synuclein. Results The mutant a-synuclein gene was successfully cloned into vector. The sequence of mutant α-synuclein was in accordance with that of wild type a-synuclein except that the number 290 base G was replaced with A. Conclusion The construction of an expression system of mutant G290A α-synuclein lays the foundation for investigating the pathogenesis of Parkinson's disease.

关 键 词：突变型 真核表达质粒 Α-SYNUCLEIN 帕金森病 脑组织 人胚 构建 N基因 酶切 野生型 

分 类 号：R742.5[医药卫生—神经病学与精神病学]