An alternative fast and convenient genotyping method for the screening of angiotensin converting enzyme gene polymorphisms.
An alternative fast and convenient genotyping method for the screening of angiotensin converting enzyme gene polymorphisms.
复制标题
用于筛选血管紧张素转换酶基因多态性的另一种快速便捷的基因分型方法。
作者：
C. Tanaka;K. Kamide;S. Takiuchi;Y. Miwa;M. Yoshii;Y. Kawano;T. Miyata
血管紧张素转换酶基因 (ACE) 内含子 16 的插入/缺失 (I/D) 多态性与血浆血管紧张素转换酶 (ACE) 水平相关，据报道，具有 DD 等位基因的个体比不具有 DD 等位基因的个体更容易患心血管疾病。传统的I/D多态性筛选基因分型方法需要聚合酶链式反应(PCR)-凝胶电泳，费时费力。在本研究中，我们评估了使用 TaqMan-PCR 基因分型来筛选 I/D 多态性作为传统方法的替代方法。我们对与 I/D 多态性连锁不平衡 (LD) 的 7 个单核苷酸多态性 (SNP) 进行了基因分型，并计算了 I/D 多态性的 LD 系数。我们发现 rs4331、rs4334 和 rs4341 三个多态性表现出最高的 LD 系数（D' = 1.000；r2 = 0.967），并且通过 TaqMan-PCR 方法对 rs4341 进行基因分型在不同基因型之间产生了最好的区分度。使用 rs4341 的 TaqMan-PCR 方法对 511 个样本进行基因分型仅需 2 小时，每次测试所需的 DNA 量仅为 6 ng。在这项研究过程中，我们在 rs4316 附近的外显子 13 中发现了一个新的额外多态性（六个氨基酸的缺失）。缺失等位基因编码睾丸 ACE，但不编码血浆 ACE。我们得出结论，通过 TaqMan-PCR 方法对 rs4341 ACE 多态性进行基因分型是直接 I/D 基因分型的一种快速、方便的替代方法。我们还得出结论，睾丸 ACE 可能表现为六个氨基酸的缺失，这可能导致该酶的有害功能。
Insertion/deletion (I/D) polymorphisms in intron 16 of the angiotensin converting enzyme gene (ACE) are associated with the plasma angiotensin converting enzyme (ACE) levels, and individuals with the DD allele have been reported to be more susceptible to cardiovascular disease than those without. The conventional genotyping method for the screening of I/D polymorphisms, which involves polymerase chain reaction (PCR)-gel electrophoresis, is laborious and time-consuming. In this study, we assessed the use of TaqMan-PCR genotyping for the screening of I/D polymorphisms as a replacement for the conventional method. We genotyped seven single nucleotide polymorphisms (SNPs) in linkage disequilibrium (LD) with the I/D polymorphisms, and calculated the LD coefficients of the I/D polymorphisms. We found that three polymorphisms, rs4331, rs4334 and rs4341, exhibited the highest LD coefficients (D' = 1.000; r2 = 0.967) and that the genotyping of rs4341 by the TaqMan-PCR method yielded the best discrimination among the different genotypes. Genotyping of 511 samples took only 2 h and the amount of DNA required for each test was only 6 ng by the TaqMan-PCR method using rs4341. In the course of this study, we identified a novel additional polymorphism (a deletion of six amino acids) in exon 13, near rs4316. The deletion allele encoded the testicular ACE, but not the plasma ACE. We concluded that genotyping of the rs4341 ACE polymorphism by the TaqMan-PCR method is a fast and convenient alternative method for direct I/D genotyping. We also concluded that testicular ACE may manifest a deletion of six amino acids that may result in deleterious function of this enzyme.
DOI：
10.1073/pnas.86.20.7741
发表时间：
1989-10-01
影响因子：
11.1
作者：
EHLERS, MRW;FOX, EA;RIORDAN, JF
通讯作者：
RIORDAN, JF
影响因子：
9.8
作者：
Zhu, XF;Bouzekri, N;Ward, R
通讯作者：
Ward, R