MOLECULAR ANALYSIS OF DELETION MUTATIONS

该提案的目的是开发方法和试剂 用于在分子水平上研究缺失突变。 等 尚未开发用于分析缺失突变的技术 这是由一类重要的环境因素引起的 诱变剂/致癌物，并且是评估潜在 这些化合物对人体的危害。 次黄嘌呤鸟嘌呤 磷酸核糖基转移酶（HPRT）基因在人类细胞中将是分子 目标 在第一阶段，反向聚合酶链反应（PCR）技术 将被应用于证明其在获得DNA序列中的用途， 断点连接和开发独特的探针（PCR和 杂交）以分析缺失位点。 此外，PCR引物- 对将从酵母人工染色体（YAC）开发， 允许测定缺失大小。 这些新的试剂和方法将用于 用于缺失突变的分子分析的II期研究 是由化学物质引起的。

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.