IN VITRO ANALYSIS OF A DNA REPLICATION APPARATUS

DNA 复制装置的体外分析

• 批准号: 3272267
• 负责人: NAVIN K SINHA
• 金额: $ 12.19万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-08-01 至 1986-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3272267
• 关键词: DNA directed DNA polymerase; bacteriophage T4; frameshift mutation; mutant; virus DNA; virus genetics

It is planned to use purified replication proteins of bacteriophage T4 to analyze the mechanisms involved in generation of high fidelity during DNA replication. The accuracy of T4 DNA polymerase (in the absence of assessory proteins) during the duplication of natural DNA templates will be measured in a number of different DNA sequence contexts. The influence of a variety of different factors, with special emphasis on polymerase accessory proteins, on the accuracy of this reaction will be measured. The role of these proteins will also be investigated by substituting mutant species in DNA replication reactions catalyzed by a seven enzyme replication complex. Attempts will be made to analyze the relative contributions of base mispairing, base stacking, efficiency of proofreading and longer range DNA sequence factors on the accuracy of DNA replication. An in vitro system for the study of frameshift mutations will be established so that the mechanisms involved in their generation can be analyzed in great detail.

计划使用T4噬菌体的纯化复制蛋白来 分析DNA过程中高保真度产生的机制 复制。T4DNA聚合酶的准确性(在缺乏 评估蛋白质)在复制天然DNA模板时将 在许多不同的DNA序列环境中测量。的影响 各种不同的因素，特别是聚合酶 辅助蛋白，对这一反应的准确性将进行测定。这个 这些蛋白质的作用也将通过取代突变体进行研究。 七种酶催化的DNA复制反应中的物种 复制复合体。将尝试分析这一亲属 碱基错配、碱基堆积、校对效率的贡献 以及更长范围的DNA序列因素影响DNA复制的准确性。 一个研究移码突变的体外系统将是 建立了与它们的产生有关的机制，可以 进行了非常详细的分析。