通过变性高效液相色谱法简单检测大的插入缺失：以ACE基因作为模型

Simple detection of large InDeLS by DHPLC: the ACE gene as a model.

作者信息

Koyama Renata Guedes, Castro Rosa M R P S, De Mello Marco Túlio, Tufik Sergio, Pedrazzoli Mario

机构信息

Departamento de Psicobiologia, Universidade Federal de São Paulo (UNIFESP), Rua Napoleão de Barros, 925/1 andar Vila Clementino, São Paulo 04024-002, SP, Brazil.

出版信息

J Biomed Biotechnol. 2008;2008:562183. doi: 10.1155/2008/562183.

DOI:10.1155/2008/562183

PMID:18475319

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2358980/

Abstract

Insertion-deletion polymorphism (InDeL) is the second most frequent type of genetic variation in the human genome. For the detection of large InDeLs, researchers usually resort to either PCR gel analysis or RFLP, but these are time consuming and dependent on human interpretation. Therefore, a more efficient method for genotyping this kind of genetic variation is needed. In this report, we describe a method that can detect large InDeLs by DHPLC (denaturating high-performance liquid chromatography) using the angiotensin-converting enzyme (ACE) gene I/D polymorphism as a model. The InDeL targeted in this study is characterized by a 288 bp Alu element insertion (I). We used DHPLC at nondenaturating conditions to analyze the PCR product with a flow through the chromatographic column under two different gradients based on the differences between D and I sequences. The analysis described is quick and easy, making this technique a suitable and efficient means for DHPLC users to screen InDeLs in genetic epidemiological studies.

摘要

插入缺失多态性（InDeL）是人类基因组中第二常见的遗传变异类型。对于大型InDeL的检测，研究人员通常采用PCR凝胶分析或限制性片段长度多态性（RFLP）方法，但这些方法耗时且依赖人工解读。因此，需要一种更有效的方法来对这类遗传变异进行基因分型。在本报告中，我们描述了一种以血管紧张素转换酶（ACE）基因的I/D多态性为模型，通过变性高效液相色谱法（DHPLC）检测大型InDeL的方法。本研究中靶向的InDeL的特征是有一个288 bp的Alu元件插入（I）。我们在非变性条件下使用DHPLC，根据D序列和I序列之间的差异，在两种不同梯度下对通过色谱柱的PCR产物进行分析。所描述的分析快速简便，使该技术成为DHPLC用户在遗传流行病学研究中筛选InDeL的合适且有效的手段。