MECHANISM AND REGULATION OF E. COLI RNA POLYMERASE

建议的研究将是一项系统的生物化学调查 大肠埃希氏菌的作用机制及其调控。RNA聚合酶。几个基因 监管系统将在体外进行研究。贯穿始终的焦点 将理解基因控制区中的DNA序列是如何 参与转录启动的调控。它的作用机制 几个转录激活因子和抑制因子的作用将是 检查过了。DNA超级卷曲对选定系统的影响也将是 调查过了。在两个噬菌体系统中，聚集性 将研究两个(或更多)相反启动子之间的转录。 建议的实验依赖于标准的生化分析 体外启动子强度的测定。这些成果将得到补充。 在某些情况下，直接测量体内RNA链的起始 频率和在其他情况下的体外结合的直接测量 RNA聚合酶和其他调节蛋白的结合。以下是问题的答案 我们提出的问题将有助于对 基因表达的控制。

The research proposed will be a systematic biochemical investigation of the mechanism and regulation of E. coli. RNA Polymerase. Several gene regulatory systems will be investigated in vitro. The focus throughout will be to understand how DNA sequences in the control regions of genes contribute to the regulation of transcription initiation. The mechanism of action of several transcriptional activators and repressors will be examined. The effects of DNA supercoiling on selected systems will also be investigated. In two bacteriophage systems, the effects of convergent transcription between two (or more) opposing promoters will be studied. The experiments proposed rely on standard biochemical assays for determining in vitro promoter strength. These results will be complemented in some cases with direct measurements of in vivo RNA chain initiation frequency and in other cases with direct measurements of in vitro binding of RNA polymerase and other regulatory proteins. The answers to the questions we have posed will contribute to the biochemical description of the control of gene expression.