Genotyping the GGGCGG tandem repeat promoter polymorphism in the 5-lipoxygenase enzyme gene (ALOX5) by pyrosequencing assay.
Genotyping the GGGCGG tandem repeat promoter polymorphism in the 5-lipoxygenase enzyme gene (ALOX5) by pyrosequencing assay.
复制标题
通过焦磷酸测序分析对 5-脂氧合酶基因 (ALOX5) 中的 GGGCGG 串联重复启动子多态性进行基因分型。
DOI：
10.1089/gtmb.2008.0103
复制
发表时间：
2009
影响因子：
1.4
通讯作者：
Langaee,TaimourY
中科院分区：
文献类型：
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作者：
Schentrup,AnzeelaM;Allayee,Hooman;Lima,JohnJ;Johnson,JulieA;Langaee,TaimourY
目的对于许多具有生物学意义的重复基因多态性，特别是在基因组富含GC的区域，有效的基因分型方法是有限的。特别是ALOX 5启动子区的短串联重复序列多态性[GGCGGG]被认为是炎症性疾病的重要标志。我们开发了一种焦磷酸测序检测基因型theALOX 5短串联重复序列多态性使用焦磷酸测序技术，这将使评估这一重要的遗传标记在大，不同的人群更容易比使用当前的methods.Materials和MethodsWe使用了巢式聚合酶链反应方法扩增DNA焦磷酸测序。在先前收集的人类DNA样本的两个队列中分别评估了188和1032个样本的群体等位基因频率。用16份已知基因型的遗传样本验证了该方法的准确性，结果与讨论基因型与已知基因型样本的符合率为100%。欧洲裔美国人、西班牙裔美国人和非洲裔美国人的基因型频率与先前发表的结果一致（野生型纯合子分别为66%、64%和19%）。这里提出的方法将有利于在大样本的种族和/或种族混合的遗传关联和药物基因组学研究的多态性。
AimsEfficient genotyping methods for many biologically significant repeat genetic polymorphisms, particularly in GC-rich regions of the genome, are limited. In particular, a short tandem repeat polymorphism [GGCGGG] in the promoter region ofALOX5has been implicated as an important marker for inflammatory diseases. We developed a pyrosequencing assay to genotype theALOX5short tandem repeat polymorphism using pyrosequencing technology that will make assessing this important genetic marker in large, diverse populations more accessible than using current methods.Materials and MethodsWe used a nested polymerase chain reaction approach to amplify DNA for pyrosequencing. Population allele frequencies were assessed in two cohorts of previously collected human DNA samples with 188 and 1032 samples, respectively. Sixteen genetic samples with known genotypes were used to confirm the accuracy of the method.Results and DiscussionGenotypes were 100% concordant with samples of known genotype. Genotype frequencies in European American, Hispanic, and African American agreed with previously published results (wild-type homozygotes 66%, 64%, and 19%, respectively). The method presented here will facilitate both genetic association and pharmacogenomic research on this polymorphism in large samples that are ethnically and/or racially admixed.