NUCLEIC ACID STRUCTURE-FUNCTION IN GENE EXPRESSION

在这项申请中提出的生化实验试图识别 具有特殊功能的核酸序列和结构的特征 作为RNA聚合酶和RNA加工酶的底物，这引发了 在RNA合成过程中终止步骤，以及区分效率 来自蛋白质生物合成的低效启动区。实验 选择允许进行体外生化分析和研究的系统 这些元素在体内的功能。函数行列式 转录终止、RNA加工和核糖体的信号 结合将通过对核苷酸的系统操作来确定 序列。单链和双链核酸酶和 化学试剂将作为RNA二级结构的实验探针 结构以获得被认为是分子内相互作用的证据 强烈影响基因表达过程中的信使核糖核酸活性。这个 核糖体对几种启动子的独立识别能力 将进行体外和体内检测。换算收益率将是 在为参考提供标准的条件下进行测量。

Biochemical experiments proposed in this application seek to identify features of nucleic acid sequence and structure that function specifically as substrates for RNA polymerase and RNA processing enzymes, that trigger the termination step during RNA synthesis, and that distinguish efficient from inefficient initiator regions for protein biosynthesis. Experimental systems are chosen to permit both in vitro biochemical analysis and studies of the in vivo functioning of these elements. The functional determinants of signals for transcription termination, RNA processing, and ribosome binding will be identified by systematic manipulation of nucleotide sequence. Nucleases specific for both single and double strands and chemical reagents will be used as experimental probes of RNA secondary structure to obtain evidence for intramolecular interactions believed to strongly influence mRNA activity during steps of gene expression. The capacity for independent recognition by ribosomes of several initiators in in vitro and in vivo assays will be examined. Translational yields will be measured under conditions that provide a standard for reference.