DETERMINATION OF MUTATIONAL SPECTRA IN THE LACI GENE

描述(改编自调查人员摘要)：申请者 建议建立一种高效的杂交筛选试验 基于微孔转移销法的LacI基因突变 和一种“探头”汇集策略。其目标是提供“突变” 指纹或光谱“能够定义水平和机制 由个别诱变化合物诱发的突变。这项测试将使用 Stratagene的BigBlue转基因系统恢复突变。然而， 目前的提议寻求制定一种战略，使荧光素- 与部分LacI突变互补的标记寡核苷酸探针 对生物素标记的PCR产物进行特定的退火热处理 链霉亲和素涂层的针脚。阳性杂交事件被揭示出来 通过使用抗荧光素-马萝卜的比色反应 微孔中的过氧化物酶结合物。 建议的商业应用：不可用

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