Mutation detection in the human HSP70B′ gene by denaturing high-performance liquid chromatography
Mutation detection in the human HSP70B′ gene by denaturing high-performance liquid chromatography
复制标题
变性高效液相色谱法检测人HSP70B′基因突变
DOI：
--
复制
发表时间：
2000
期刊：
影响因子：
--
通讯作者：
S. Calderwood
中科院分区：
文献类型：
--
作者：
K. Hecker;A. Asea;Kaoru Kobayashi;S. Green;D. Tang;S. Calderwood
摘要：个体基因组序列中的变异，特别是单核苷酸多态性 (SNP)，是了解基因功能的主要关键，因为它与疾病、环境影响和治疗的易感性差异有关。在本报告中，HSP70B' 基因是使用温度调制异源双链分析 (TMHA) 对在丹纳法伯癌症研究所接受热疗和放射治疗的人类前列腺癌患者的活检样本中进行突变检测的目标序列。讨论了使用 DHPLC 技术进行 TMHA 突变检测的基本原理。详细介绍了 DHPLC 突变分析的扩增子设计过程。使用 WAVEMAKERTM 软件对目标基因的完整编码序列的解链行为进行表征。根据软件预测的靶序列熔解曲线，鉴定了四个重叠的扩增子，它们跨越了 HSP70B' 基因的完整编码区，适合通过 DHPLC 进行突变检测。在 WAVE® 核酸片段分析系统上对 HSP70B' 基因的各个扩增子的 PCR 产物进行 TMHA。讨论了通过比较野生型和突变体色谱模式进行突变识别的标准。
Abstract Variances, particularly single nucleotide polymorphisms (SNP), in the genomic sequence of individuals are the primary key to understanding gene function as it relates to differences in the susceptibility to disease, environmental influences, and therapy. In this report, the HSP70B′ gene is the target sequence for mutation detection in biopsy samples from human prostate cancer patients undergoing combined hyperthermia and radiation therapy at the Dana-Farber Cancer Institute, using temperature-modulated heteroduplex analysis (TMHA). The underlying principles of TMHA for mutation detection using DHPLC technology are discussed. The procedures involved in amplicon design for mutation analysis by DHPLC are detailed. The melting behavior of the complete coding sequence of the target gene is characterized using WAVEMAKERTM software. Four overlapping amplicons, which span the complete coding region of the HSP70B′ gene, amenable to mutation detection by DHPLC were identified based on the software-predicted melting profile of the target sequence. TMHA was performed on PCR products of individual amplicons of the HSP70B′ gene on the WAVE® Nucleic Acid Fragment Analysis System. The criteria for mutation calling by comparing wild-type and mutant chromatographic patterns are discussed.
犬遗传性蜡样质脂褐质沉着症：肉碱生物合成途径缺陷的证据。
DOI：
10.1002/ajmg.1320570231
发表时间：
1995
期刊：
American journal of medical genetics
影响因子：
--
作者：
Katz,ML;Siakotos,AN
通讯作者：
Siakotos,AN
影响因子：
14.9
作者：
Liu, WG;Smith, DI;James, CD
通讯作者：
James, CD
用变性高效液相色谱法检测大量 Y 染色体双倍型多态性
影响因子：
7
作者：
Underhill, PA;Jin, L;Oefner, PJ
通讯作者：
Oefner, PJ