Koch Werner, Ehrenhaft Angela, Griesser Korinna, Pfeufer Arne, Müller Jakob, Schömig Albert, Kastrati Adnan
Deutsches Herzzentrum München and 1. Medizinische Klinik, München, Germany.
Clin Chem Lab Med. 2002 Nov;40(11):1123-31. doi: 10.1515/CCLM.2002.197.
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 (epsilon2; 112Cys/158Cys) = 7.3%, 334T/472C (epsilon3; 112Cys/158Arg) = 78.4%, and 334C/472C (epsilon4; 112Arg/158Arg) = 14.2%, respectively. ApoE genotypes were distributed as: epsilon2epsilon2 = 0.9%, epsilon2epsilon3 = 11.2%, epsilon2epsilon4 = 1.6%, epsilon3epsilon3 = 61.3%, epsilon3epsilon4 = 23.1%, and epsilon4epsilon4 = 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.
载脂蛋白E基因的多态性与外周和冠状动脉疾病以及神经退行性疾病（如散发性和晚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，用于检测位于载脂蛋白E基因启动子区的单核苷酸多态性-219G/T、内含子1转录增强子元件中的113G/C、决定成熟载脂蛋白E第112位氨基酸残基为半胱氨酸或精氨酸的334T/C以及决定第158位残基为精氨酸或半胱氨酸的472C/T。通过限制性内切酶分析证明了TaqMan系统进行基因型测定的准确性。我们测定了2349名研究对象的载脂蛋白E多态性基因型。基因型分布如下：-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%，472TT = 0.9%（p = 0.004）（括号内为哈迪-温伯格平衡估计值）。定义载脂蛋白E三种主要异构体即apoE2、apoE3和apoE4的等位基因组合的等位基因频率如下：334T/472T（ε2；112位半胱氨酸/158位半胱氨酸）= 7.3%，334T/472C（ε3；112位半胱氨酸/158位精氨酸）= 78.4%，334C/472C（ε4；112位精氨酸/158位精氨酸）= 14.2%。载脂蛋白E基因型分布如下：ε2ε2 = 0.9%，ε2ε3 = 11.2%，ε2ε4 = 1.6%，ε3ε3 = 61.3%，ε3ε4 = 23.1%，ε4ε4 = 1.9%（p = 0.014）。TaqMan检测法能够实现快速且灵敏的基因分型，尤其适用于包含大量参与者的研究。
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