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（改编自研究者的摘要）：申请人 建议开发一种有效的杂交筛选试验 基于微孔转移针方法的 lacI 基因突变 以及“探测”池策略。 目标是提供“突变 指纹或光谱”能够定义水平和机制 由单个诱变化合物引起的突变。 测试将使用 Stratagene 的 BigBlue 转基因系统可恢复突变体。 然而， 本提案旨在制定一项战略，使荧光素- 与选定的 lacI 突变互补的标记寡核苷酸探针 与固定在上的生物素标记的 PCR 产物特异性退火 链霉亲和素涂层的针。 揭示了阳性杂交事件 通过使用抗荧光素辣根的比色反应 微孔中的过氧化物酶缀合物。 拟议的商业应用： 不可用