MOLECULAR ANALYSIS OF DELETION MUTATIONS

这项建议的目标是开发方法和试剂 用于在分子水平上研究缺失突变。是这样的 还没有开发出用于分析缺失突变的技术 它们是由一类重要的环境因素引起的 诱变剂/致癌物，是评估潜力所必需的 这些化合物对人类的危害。次黄嘌呤鸟嘌呤 人类细胞中的磷酸核糖基转移酶(HPRT)基因将是 目标。在第一阶段，反向聚合酶链式反应(PCR)技术 将被应用于展示其在获取DNA序列中的应用 断点连接和开发独特的探针(包括PCR和 杂交)，用于缺失位点的分析。此外，聚合酶链式反应-- 配对将从酵母人工染色体(YAC)中开发出来，这将 允许分析删除大小。这些新的试剂和方法将被使用 在缺失突变的分子分析的第二阶段研究中 由化学试剂引起的。

The objective of this proposal is the development of methods and reagents for use in the study of deletion mutations at the molecular level. Such technology has not been developed for the analysis of deletion mutations which are induced by an important class of environmental mutagens/carcinogens and is necessary for the assessment of potential hazard to man from these compounds. the hypoxanthine guanine phosphoribosyltransferase (hprt) gene in human cells will be the molecular target. During Phase I, inverse polymerase chain reaction (PCR) technology will be applied to demonstrate its use in obtaining DNA sequence of breakpoint junctions and in developing unique probes (both PCR and hybridization) for analysis of deletion sites. In addition, PCR primer- pairs will be developed from yeast artificial chromosomes (YACs) which will allow assay of deletion size. These new reagents and methods will be used in Phase II studies for the molecular analysis of deletion mutations induced by chemical agents.