TaqMan Systems for Genotyping of Disease-Related Polymorphisms Present in the Gene Encoding Apolipoprotein E
TaqMan Systems for Genotyping of Disease-Related Polymorphisms Present in the Gene Encoding Apolipoprotein E
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DOI：
10.1515/cclm.2002.197
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发表时间：
2002-12-01
影响因子：
6.8
通讯作者：
Kastrati, Adnan
中科院分区：
文献类型：
--
作者：
Koch, Werner;Ehrenhaft, Angela;Kastrati, Adnan
编码载脂蛋白E的基因多态性与外周动脉和冠状动脉疾病以及神经退行性疾病（例如散发性和迟发性家族性阿尔茨海默病）的发病机制有关。我们开发了 TaqMan 检测系统，用于检测单核苷酸多态性 -219G/T（位于载脂蛋白 E 基因的启动子中）、113G/C（存在于内含子 1 的转录增强子元件中）、334T/C（确定 Cys 或 Arg 为成熟载脂蛋白 E 的氨基酸残基 112）和 472C/T（确定 Arg 或 Cys 为残基 158）。通过限制性内切酶分析证明了 TaqMan 系统的基因型测定。我们确定了 2349 名研究对象的载脂蛋白 E 多态性基因型。基因型分布为：-219GG= 27.3%、-219GT= 49.1%、-219TT=23.6% (p=0.435)； 113GG=41.3%、113GC=45.2%、113CC=13.5%（p=0.343）； 334TT=73.4%、334TC=24.7%、334CC=1.9%（p=0.539）； 472CC=86.3%、472CT=12.8% 和 472TT= 0.9% (p=0.004)（Hardy-Weinberg 平衡估计值在括号中给出）。定义载脂蛋白E的三种主要亚型，即apoE2、apoE3和apoE4的等位基因组合具有以下等位基因频率：334T/472T(ε2；112Cys/158Cys)=7.3%，334T/472C(ε3；112Cys/158Cys)=7.3%，334T/472C(ε3；112Cys/158Cys)=7.3%。 112Cys/158Arg)=78.4%，334C/472C(ε4；112Arg/158Arg)=14.2%。 ApoE基因型分布为：ε2ε2=0.9%，ε2ε3=11.2%，ε2ε4=1.6%，ε3ε3=61.3%，ε3ε4=23.19 1 0，ε4ε4=1.9% （p=0.014）。 TaqMan 检测可实现快速、灵敏的基因分型，特别适合包含大量参与者的研究。
Polymorphisms of the gene encoding apolipoprotein E have been implicated in the pathogenesis of peripheral and coronary artery disease and neurodegenerative disorders such as sporadic and late-onset familial forms of Alzheimer's disease. We have developed TaqMan assay systems for the single nucleotide polymorphisms -219G/T, located in the promoter of the apolipoprotein E gene, 113G/C, present in the transcriptional enhancer element of intron 1, 334T/C, determining Cys or Arg as amino acid residue 112 of mature apolipoprotein E, and 472C/T, determining Arg or Cys as residue 158. The accuracy of genotype determination with the TaqMan systems was demonstrated by analyses with restriction endonucleases. We determined the genotypes of the apolipoprotein E polymorphisms in 2349 study subjects. The genotypes were distributed as: -219GG= 27.3%, -219GT= 49.1%, and -219TT=23.6% (p=0.435); 113GG=41.3%, 113GC= 45.2%, and 113CC=13.5% (p=0.343); 334TT=73.4%, 334TC=24.7%, and 334CC= 1.9% (p=0.539); 472CC=86.3%, 472CT=12.8%, and 472TT= 0.9% (p=0.004) (Hardy-Weinberg equilibrium estimates are given in parentheses). The allele combinations which define the three major isoforms of apolipoprotein E, namely apoE2, apoE3, and apoE4, had the following allele frequencies: 334T/472T (epsilon 2; 112Cys/158Cys)=7.3%, 334T/472C (epsilon 3; 112Cys/158Arg)=78.4%, and 334C/472C (epsilon 4; 112Arg/158Arg)=14.2%, respectively. ApoE genotypes were distributed as: epsilon 2 epsilon 2=0.9%, epsilon 2 epsilon 3= 11.2%, epsilon 2 epsilon 4= 1.6%, epsilon 3 epsilon 3=61.3%, epsilon 3 epsilon 4=23.19 1 0, and epsilon 4 epsilon 4=1.9% (p=0.014). The TaqMan assays allow for fast and sensitive genotyping and are especially suitable for studies including large numbers of participants.