DNA--MEMBRANE INTERACTIONS

DNA--膜相互作用

• 批准号: 2734469
• 负责人: IAN J MOLINEUX
• 金额: $ 22.45万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-04-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2734469
• 关键词: DNA binding protein; Escherichia coli; bacteriophage T7; cell membrane; gene mutation; genetic mapping; laboratory rabbit; methylation; molecular cloning; protein structure function; virus DNA; virus infection mechanism

DESCRIPTION: This project is a comprehensive study, using primarily genetic and molecular genetic methods, of the process of entry of phage T7 DNA into infected cells. The PI has developed an assay, based on the time of methylation of GATC sequences (that have been placed at specific locations on phage DNA) by dam methylase. Under normal conditions about 1 kb of phage DNA enters and is transcribed before the remainder of the DNA enters the cytoplasm. Both cis- and trans-acting phage mutants will be obtained to identify DNA sites and phage proteins that participate in this process. E. coli mutants, that absorb T7 but are not killed, will be isolated to identify potential host components of the system. The phage proteins that enter the cell with DNA will be identified and the initial locations of both the DNA and the proteins will be determined. Initial attempts will be made study the binding of phage proteins to the leading end of phage DNA. Other objectives are a preliminary analysis of the energetics of DNA translocation and characterization of the sequence on phage T5 that is responsible for arrest of translocation of T5 DNA.

描述：该项目是一项全面的研究，主要使用遗传 和分子遗传学方法，噬菌体T7 DNA进入的过程 感染细胞。 PI根据时间开发了一种测定法 GATC序列的甲基化（已放置在特定位置 在噬菌体DNA上）通过大坝甲基化合酶。 在正常条件下约1 kb的噬菌体 DNA进入并在其余DNA进入之前转录 细胞质。 顺式和反式噬菌体突变体将获得 确定参与此过程的DNA位点和噬菌体蛋白。 E. 吸收T7但未杀死的大肠杆菌突变体将被隔离为 确定系统的潜在主机组件。 噬菌体蛋白 将确定使用DNA输入细胞，并且两者的初始位置 将确定DNA和蛋白质。 将进行初步尝试 研究噬菌体蛋白与噬菌体DNA的铅端的结合。 其他 目标是对DNA易位能量学的初步分析 以及负责噬菌体T5序列的表征 T5 DNA易位的停滞。