DETERMINATION OF MUTATIONAL SPECTRA IN THE LACI GENE

LACI 基因突变谱的测定

• 批准号: 2852955
• 负责人: JACK R. U'REN
• 金额: $ 10万
• 依托单位: SAIGENE CORPORATION
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-02-08 至 1999-08-07
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2852955
• 关键词: fluorescent dye /probe; gene expression; gene mutation; genetic screening; mutagens; nucleic acid hybridization; nucleic acid probes; nucleic acid sequence; oligonucleotides; restriction fragment length polymorphism; technology /technique development

DESCRIPTION (Adapted from investigator's abstract): The applicants propose development of an efficient hybridization screening test for mutations in the lacI gene based on a microwell transfer pin approach and a "probe" pooling strategy. The goal is to provide "mutational fingerprints or spectra" capable of defining levels and mechanism of mutation induced by individual mutagenic compounds. The test will use Stratagene's BigBlue transgenic system to recover mutants. However, the present proposal seeks to develop a strategy whereby fluorescein- labeled oligonucleotides probes complementary to selected lacI mutations are specifically annealed to biotin-labeled PCR products immobilized on streptavidin-coated pins. Positive hybridization events are revealed by a colorimetric reaction using anti-fluorescein-horse radish peroxidase conjugates in microwells. PROPOSED COMMERCIAL APPLICATION: Not available

描述（根据调查员的摘要改编）：申请人 提出开发有效的杂交筛选测试 基于Microwell转移销方法的LACI基因中的突变 以及“探测”汇总策略。 目标是提供“突变 指纹或光谱“能够定义水平和机制 由个体诱变化合物诱导的突变。 测试将使用 Stratagene的BigBlue转基因系统以恢复突变体。 但是， 目前的建议旨在制定一项策略，荧光素 - 标记的寡核苷酸探针互补与选定的LACI突变 专门将固定在生物素标记的PCR产物中退火 链霉亲和素涂层的销钉。 揭示了积极的杂交事件 通过使用抗荧光素 - 马萝卜的比色反应 过氧化物酶偶联物中的微孔。 拟议的商业应用程序：不可用