[Detection of single-nucleotide polymorphisms in insulin receptor substrate-2 gene 3'-untranslated region by denaturing high-performance liquid chromatography].

Hunan yi ke da xue xue bao = Hunan yike daxue xuebao = Bulletin of Hunan Medical University Pub Date : 2003-08-01

Wei-min Zeng, Shu-hua Chen, Ping Xie

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Abstract

Objective: To explore the use of denaturing high-performance liquid chromatography (DHPLC) in detecting single-nucleotide polymorphisms(SNPs) of insulin receptor substrate-2(IRS-2) gene 3'-untranslated region (3'-UTR).

Methods: We detected the SNPs and mutation of IRS-2 gene 3'-UTR sequence, with polymerase chain reaction (PCR), DHPLC, single-strand conformation polymorphism (SSCP), and DNA sequence analysis respectively in 30 Type 2 diabetic subjects and 30 healthy controls.

Results: The SNPs of IRS-2 gene 3'-UTR in 4 patients with Type 2 diabetes mellitus were detected by PCR-DHPLC and were identified by DNA sequencing.

Conclusion: DHPLC is a rapid and automated technology, which is simpler and more accurate for detecting longer fragment of DNA than SSCP. The present method lays a foundation for further studying the relationship between the mutation of IRS-2 gene 3'-UTR and Type 2 diabetes mellitus.

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【变性高效液相色谱法检测胰岛素受体底物2基因3′-非翻译区单核苷酸多态性】。

目的:探讨变性高效液相色谱(DHPLC)检测胰岛素受体底物-2(IRS-2)基因3′-非翻译区(3′-UTR)单核苷酸多态性(snp)的方法。方法:采用聚合酶链反应(PCR)、DHPLC、单链构象多态性(SSCP)和DNA序列分析，分别检测30例2型糖尿病患者和30例健康对照的IRS-2基因3′-UTR序列的snp和突变。结果:采用PCR-DHPLC检测4例2型糖尿病患者IRS-2基因3′-UTR的snp，并进行DNA测序鉴定。结论:DHPLC是一种快速、自动化的检测技术，与SSCP相比，DHPLC检测长片段DNA更简单、准确。本方法为进一步研究IRS-2基因3′-UTR突变与2型糖尿病的关系奠定了基础。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Hunan yi ke da xue xue bao = Hunan yike daxue xuebao = Bulletin of Hunan Medical University

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