Genotyping the GGGCGG Tandem Repeat Promoter Polymorphism in the 5-Lipoxygenase Enzyme Gene (ALOX5) by Pyrosequencing Assay

Anzeela M. Schentrup; Hooman Allayee; John J. LimaJulie A. JohnsonTaimour Y. Langaee

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Genotyping the GGGCGG Tandem Repeat Promoter Polymorphism in the 5-Lipoxygenase Enzyme Gene (ALOX5) by Pyrosequencing Assay

机译：基因分型的GGGCGG串联重复启动子5-Lipoxygenase酶基因多态性(ALOX5)通过焦磷酸测序分析

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Efficient 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 of ALOX5 has been implicated as an important marker for inflammatory diseases. We developed a pyrosequencing assay to genotype the ALOX5 short 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 Methods: We 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 Discussion: Genotypes 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.

机译：许多生物高效的基因分型方法重要的重复基因多态性,尤其是GC-rich区域的基因组,是有限的。在启动子多态性(GGCGGG)地区ALOX5被牵连是一个重要的标志炎性疾病。焦磷酸测序鉴定基因型ALOX5短串联重复序列多态性通过焦磷酸测序技术,将评估重要的遗传标记在很大,多样化比使用当前的人口更容易方法。放大DNA聚合酶链反应方法焦磷酸测序。在两群之前进行评估吗收集了188年和1032年人类DNA样本样本,分别。与已知的基因型是用来确认的该方法的准确性。基因型是100%和样品一致已知的基因型。美国人,西班牙裔,非洲裔美国人同意了与以前公布的结果(野生型比如66%、64%和19%,分别)。方法将促进遗传协会和药物基因组学的研究在这个大样本的多态性种族和/或种族混血。

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《Genetic testing and molecular biomarkers》 |2009年第3期|361-365|共5页
作者
Anzeela M. Schentrup; Hooman Allayee; John J. LimaJulie A. JohnsonTaimour Y. Langaee;
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作者单位

Center for Pharmacogenomics, College of Pharmacy, University of Florida, Gainesville, Florida.;

Centers for Clinical Pediatric Pharmacology and Pharmacogenetics, Nemours Children's Clinic, Jacksonville, Florida.;

Keck School of Medicine, University of Southern California, Los Angeles, California.;

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正文语种英语
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ALOX5 gene; promoter regions; genetic associationEnzyme GeneGenotypepolymorphismTandem Repeat SequencespyrosequencingNested Polymerase Chain ReactionMicrosatellite RepeatsGenetic MarkersArachidonate 5-LipoxygenaseGenotyping;

机译：ALOX5基因、启动子区域基因重复SequencespyrosequencingNested聚合酶链ReactionMicrosatellite RepeatsGeneticMarkersArachidonate 5-LipoxygenaseGenotyping;

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Genotyping the GGGCGG Tandem Repeat Promoter Polymorphism in the 5-Lipoxygenase Enzyme Gene (ALOX5) by Pyrosequencing Assay

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