LARGE SCALE IDENTIFICATION OF DNA VARIANTS

拟议的项目旨在开发识别 高密度寡核苷酸芯片在人类DNA变异研究中的应用 （非正式地称为DNA芯片）。 重点发展 两个关键应用的技术。 1.单核苷酸多态性（SNP）。我们将： (i)开发可靠的方法来识别单核苷酸 多态性（SNP）; (ii)通过识别STS中的SNP来测试方法 覆盖约1.5Mb的基因组DNA; (iii)构建第三代人类基因组遗传图谱， 2000个SNP，杂合性约为40%，已知图谱 （四）开发一个强大的基因分型系统，以使用这第三个- 生成遗传图谱，包括寡核苷酸阵列和高效 用于多重扩增的实验室方案。 2.常见的基因变异。 一些基因中的常见变异 (such如ApoE、FactorV、CCR-5和MTHFR）最近已被证明 与疾病易感性有关。系统地应用 这种强有力的人类遗传学方法将需要：（a）人口 调查，以确定所有常见的变异基因涉及在一个给定的 （B）关联性研究，以将 常见的变异与疾病风险。 我们将专注于第一项任务。 我们将： (i)探索执行人口调查的有效方法， 通过使用基因组DNA或mRNA，编码序列的变异。 (ii)应用这些技术来识别基因中的常见变异 涉及两个生理系统--（a）血液凝固和（B） 脂质运输和代谢--已经知道是一些 与疾病风险有重要关系的常见变异。

The proposed project aims to develop technology for identification of human DNA variation, by using high-density oligonucleotide arrays (informally referred to as DNA chips). We will focus on developing technology for two key applications. 1. Single Nucleotide Polymorphisms (SNPs). We will: (i) Develop robust methods to identify the single nucleotide polymorphisms (SNPs); (ii) Test the methods by identifying SNPs in STSs covering approximately 1.5 Mb of genomic DNA; (iii) Construct a third-generation genetic map of the human genome with 2000 SNPs, with heterozygosity approximately 40 percent and known map locations; (iv) Develop a robust genotyping system for using this third- generation genetic map, including an oligonucleotide array and efficient laboratory protocols for multiplex amplification. 2. Common Variation in Genes. Common variants in a number of genes (such as ApoE, FactorV, CCR-5, and MTHFR) have recently been shown to be associated with disease susceptibilities. Systematic application of this powerful approach to human genetics will require: (a) population surveys to identify all common variants in the genes involved in a given physiological system; and (b) association studies to correlate the common variants with disease risk. We will focus on the first task. We will: (i) Explore efficient ways to perform a population survey to discover variation in coding sequences, either by using genomic DNA or mRNA. (ii) Apply the techniques to identify the common variants in the genes involved in two physiological systems--(a) blood coagulation and (b) lipid transport and metabolism--already known to be the site of some common variants having important relationships to disease risk.