PRO FLUORESCENT NUCLEOTIDE ANALOGS FOR GENETIC ANALYSIS

用于遗传分析的专业荧光核苷酸类似物

• 批准号: 2652131
• 负责人: MICHAEL T BOYCE-JACINO
• 金额: $ 2.6万
• 依托单位: ORCHID BIOCOMPUTER, INC.
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-05-15 至 1998-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2652131
• 关键词: chemical synthesis; fluorescent dye /probe; gene mutation; genetic techniques; method development; molecular genetics; nucleotide analog

DESCRIPTION: (adapted from applicant's abstract) Nucleotide analogs labeled with detectable moieties, such as fluorescent dyes, are widely used in research and diagnostic assays. Chain terminating nucleotide analogs (dideoxynucleotides) are especially useful in DNA sequencing applications and primer extension assays, such as minisequencing or genetic bit analysis (GBA). They propose to develop a unique set of pro-fluorescent dideoxynucleotide analogs that will enable more rapid and sensitive mutation detection as well a provide unique substrates for polymerase activity assays, such as for cell viability. The proposed analogs are non-fluorescent as free nucleotide, however, upon incorporation into DNA by a DNA polymerase, a fluorophore is released into solution. The amount of fluorescence released into solution is directly correlated with polymerase activity, thus providing a real-time assay for incorporation of a specific base analog. They propose to synthesize and test the concept of pro-fluorescent dideoxynucleotide analogs during Phase I of this grant and if successful, develop all four base analogs and assay conditions during Phase II. Successful completion of this project will result in a series of pro-fluorescent nucleotide analogs with broad ranging utility in genetic and biochemical research and diagnostics. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE

描述：（改编自申请人的摘要）核苷酸类似物 标记有可检测的部分，例如荧光染料， 用于研究和诊断分析。 链终止核苷酸 类似物（双脱氧核苷酸）在DNA测序中特别有用 应用和引物延伸测定，如微测序或 遗传位分析（GBA）。 他们建议开发一套独特的 前荧光双脱氧核苷酸类似物， 和敏感的突变检测，以及提供独特的底物， 聚合酶活性测定，例如用于细胞活力。 拟议 类似物作为游离核苷酸是非荧光的，然而， 通过DNA聚合酶掺入DNA，释放荧光团 变成溶液。 释放到溶液中的荧光量为 与聚合酶活性直接相关，因此提供了实时 用于掺入特定碱基类似物的测定。 他们建议合成和测试前荧光的概念， 双脱氧核苷酸类似物在第一阶段，如果 成功地开发了所有四种碱基类似物和测定条件， 第二阶段。这个项目的成功完成将导致一系列 具有广泛用途的前荧光核苷酸类似物， 基因和生化研究和诊断。 拟议商业应用：不可用