疾病基因克隆的策略及主要方法

疾病基因克隆的策略及主要方法申海鹰 周元国（第三军医大学野战外科研究所分子生物学中心，重庆400042） 摘要 疾病基因的分离和克隆是功能基因组学的研究热点，具体策略的选择取决于疾病背景资料的掌握

疾病基因克隆的策略及主要方法 申海鹰 周元国（第三军医大学野战外科研究所分子生物学中心，重庆 400042） 摘要 疾病基因的分离和克隆是功能基因组学的研究热点，具体策略的选择取决于 疾病背景资料的掌握程度，为能快速、准确地克隆出目的基因，本文介绍两类常用 的基因克隆策略——定位克隆策略、功能克隆策略——及其主要方法，如：家系连 锁分析、等位基因共占法、人群相关分析法、抑制性消减杂交、差示反转录PCR、 差异消减显示法、代表性差异分析法、比较基因组杂交等，并作简要的评价。 关键词 基因；定位克隆；消减杂交 学科分类号 Q785 Strategies and methods for cloning pathogenic gene SHEN Hai-ying, ZHOU Yuan-guo. (Center of Molecular Biology, Research Institute of Surgery and Daping Hospital, Third Military Medical University, Chongqing 400042) Abstract Isolation and cloning of pathogenic gene is ahot spot in functional genome study, while it is the disease background which decides the selection of the strategies. Two strategies, mapping strategy and functional cloning strategy, which can clone the objective gene rapidly and accurately were introduced. Some main methods including family-based linkage analysis, allele sharing method, population association analysis, suppression subtractive hybridization (SSH), differential display reverse-transcription PCR (DD-RT-PCR), differential subtraction display (DSD), representational difference analysis (RDA),