CHARACTERIZATION OF THE HUMAN GENOME

描述(改编自《调查者摘要》)：DNA研究 熔融的提出将导致一种新的综合方法 人类基因中序列变异的检测。方法不同于 目前的程序提供快速、易于自动化、可靠和 完全详尽的多态和突变的迹象。这种方法， 就像变性梯度凝胶电泳法一样，依赖于融化 关于DNA熔化温度的理论和微扰 存在单碱基(或更大的)序列变化或错配；但 荧光取代了电泳法。在冰冻检测中 电泳法，这项技术将允许快速、同时检查 许多小样本，每个样本都有相当长的序列。该项目 将包括以下工作：(1)引入荧光作为 DNA融化的指示剂；(2)与A结合的DNA的熔融观察 二氧化硅表面；(3)热稳定性的分布分析 使用大量突变的微生物基因；(4)严格测试融化 (5)将该检测系统应用于汽车尾气检测中 人类Y染色体基因突变(与D.Page合作)；和(6) DNA结构域结构与组分的相关性分析 它的环境-离子等，当被并入后续工程时 开发方面，可以预期这种方法将对 大规模突变分析和筛选的可行性。

DESCRIPTION (Adapted from the Investigator's Abstract): Studies on DNA melting are proposed that will lead to a novel method for comprehensive detection of sequence variants in human genes. The method differs from current procedures in offering rapid, readily automatable, reliable, and fully exhaustive indication of polymorphisms and mutations. The approach, like that of denaturing gradient gel electrophoresis, depends on melting theory and on the perturbation of the melting temperature of DNA by the presence of single-base (or larger) sequence changes or mismatches; but electrophoresis is replaced by fluorescence. In freezing detection from electrophoresis, the technique will permit rapid, simultaneous scrutiny of many small samples, a substantial length of sequence in each. The project will consist of the following efforts: (1) introduction of fluorescence as an indicator of DNA melting; (2) melting observation of DNA bound to a silica surface; (3) analysis of the distribution of thermal stability in a microbial gene using a large set of mutants; (4) rigorously testing melting theory with those results; (5) applying the detection system to detection of mutants in human Y chromosome genes (in collaboration with D. Page); and (6) analysis of the dependence of the domain structure of DNA on components of its environment - ions, etc. When incorporated into subsequent engineering development, this approach can be expected to effect a major change in the feasibility of large-scale mutation analysis and screening.