PCR-RFLP genotyping assay for the Bcl I polymorphism of the beta-fibrinogen gene.
PCR-RFLP genotyping assay for the Bcl I polymorphism of the beta-fibrinogen gene.
复制标题
β-纤维蛋白原基因 Bcl I 多态性的 PCR-RFLP 基因分型测定。
DOI：
10.1089/109065701750168707
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
Booyse,FM
中科院分区：
文献类型：
--
作者：
Sell,SM;Song,C;Booyse,FM
β-纤维蛋白原基因的多态性已被证明影响血浆纤维蛋白原水平和冠状动脉疾病（CAD）的风险。我们有兴趣开发一种自动化的，基于PCR的基因分型 本研究旨在探讨冠心病及其相关的主动脉僵硬度与β-纤维蛋白原基因的BclI等位基因之间的关系。我们建立了一种快速检测β-纤维蛋白原基因BclI多态性的PCR-RFLP方法。我们进行了基因组DNA的直接PCR，以促进β-纤维蛋白原基因侧翼区的测序。使用这个新的序列信息， 设计了与Bcl I多态性位点相邻的引物。其中一个引物用荧光团标记，以便于检测片段。DNA片段分析 使用自动毛细管电泳仪（ABI 310）。我们建立了一种改进的PCR-RFLP高通量半自动检测Bcl I基因多态性的方法 β-纤维蛋白原基因。本试验将支持人群研究所需的大样本量筛选。
Polymorphisms of the β-fibrinogen gene have been shown to affect plasma fibrinogen levels and risk of coronary artery disease (CAD). We were interested in developing an automated, PCR-based genotyping assay for the purpose of exploring relationships between CAD and CAD-associated aortic stiffness and theBclI allele of the β-fibrinogen gene. We have developed a rapid PCR-RFLP assay for theBclI polymorphism of the β-fibrinogen gene. We carried out direct PCR of genomic DNA to facilitate sequencing of the flanking region of the β-fibrinogen gene. Using this new sequence information, primers were designed which border the site of theBclI polymorphism. One of the primers was labeled with a fluorophore to facilitate detection of the fragments. DNA fragment analysis was carried out using an automated capillary electrophoresis instrument (ABI310). We have developed an improved PCR-RFLP high-sample-throughput assay for the semiautomated detection of theBclI polymorphism of the β-fibrinogen gene. This assay will support screening of large sample sizes required for population studies.