CHARACTERIZATION OF THE HUMAN GENOME

人类基因组的特征

• 批准号: 2519124
• 负责人: LEONARD S LERMAN
• 金额: $ 35.64万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-09-01 至 1999-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2519124
• 关键词: DNA; biological polymorphism; chromosome aberrations; computer assisted sequence analysis; diagnosis design /evaluation; fluorescent dye /probe; genetic mapping; genetic techniques; genome; human genetic material tag; image processing; nucleic acid denaturation; nucleic acid probes; nucleic acid sequence; nucleobase; polymerase chain reaction; thermostability

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.

描述（改编自研究者摘要）：DNA研究 熔化的建议，将导致一种新的方法，全面 检测人类基因中的序列变异。 该方法不同于 目前的程序提供快速，易于自动化，可靠， 多态性和突变的完全详尽的指示。 这种做法， 类似于变性梯度凝胶电泳， 理论和DNA解链温度的扰动 存在单碱基（或更大）序列变化或错配;但 电泳被荧光取代。 在冷冻检测中， 电泳，该技术将允许快速，同时审查 许多小样本，每个样本中有相当长的序列。 项目 将包括以下努力：（1）引入荧光作为 DNA解链的指示剂;（2）结合到DNA上的DNA的解链观察， （3）分析了硅表面的热稳定性分布， 使用大量突变体的微生物基因;（4）严格测试融化 （5）将该检测系统应用于检测 人类Y染色体基因的突变体（与D.（6）;（6） 分析DNA结构域的组成成分依赖性 其环境-离子等，当纳入后续工程 发展，这一做法可以预期将影响重大变化， 大规模突变分析和筛选的可行性。