REGULATION OF EXOGENOUSLY INDUCED OPERON

外源诱导操纵子的调控

• 批准号: 3280184
• 负责人: JEN SHIANG HONG
• 金额: $ 15.62万
• 依托单位: BOSTON BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-04-01 至 1989-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3280184
• 关键词: Escherichia coli; Salmonella typhimurium; bacterial genetics; biological transport; crosslink; gene expression; genetic manipulation; genetic transcription; genetic translation; immunochemistry; molecular cloning; mutant; nucleic acid sequence; operon; phosphoglycerate; radiotracer; regulatory gene

The genetic expression of the phosphoglycerate transport system of S. typhimurium is induced by external inducers but not by internal inducers. The long-term objectives of the proposed research are to gentically and biochemically characterize the regulatory machinery involved in gene expression of this transport system and to understand the biochemical and molecular mechanism underlying this novel gene regulation. During the current grant period we dissected the genetic regulatory machinery, determined the DNA sequence, identified gene products and determined the cellular locations of these products. The specific aims of the present proposal are: to isolate, purify and characterize the pgt regulatory proteins; to study the regulatory mechanism in in vitro transcription-translation coupled system in order to determine the functional role of each regulatory proteins with respect to inducer binding, DNA binding and signal transmission; to analyze pgt transcripts and determine pgtP transcription initiation start and DNA binding regions, in order to distinguish two models of regulation and to define the control regions; to isolate constitutive regulatory mutants by genetic selection and site-directed mutagenesis and to characterize the mutant proteins; to study processing of regulatoryp proteins and determine the cleavage sites; and to perform chemical cross-linking experiments to investigate the interaction of the regulatory proteins.

对S. 鼠伤寒沙门氏菌是由外部诱导剂诱导的，而不是由内部诱导剂诱导的。 拟议研究的长期目标是从基因和 从生物化学上描述基因调控机制 表达这种运输系统，并了解生物化学和 这种新的基因调控的分子机制。 期间 在当前的资助期内，我们剖析了基因调控机制， 确定了DNA序列，鉴定了基因产物， 这些产品的手机定位。 目前的具体目标 建议：分离，纯化和表征PGT调节 蛋白质;研究体外调控机制 转录-翻译偶联系统，以确定 每种调节蛋白对诱导物的功能作用 结合、DNA结合和信号传递;分析pgt转录物 并确定pgtP转录起始和DNA结合区域， 为了区分两种监管模式， 区域;通过遗传选择分离组成型调节突变体 和定点诱变，并表征突变蛋白; 研究调节型蛋白的加工过程并确定切割位点; 并进行化学交联实验，以研究 调节蛋白的相互作用。