REGULATION OF HISTIDINE BIOSYNTHESIS IN YEAST

酵母中组氨酸生物合成的调控

• 批准号: 3287064
• 负责人: GERALD R FINK
• 金额: $ 31.55万
• 依托单位: WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-07-01 至 1985-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3287064
• 关键词: Saccharomyces; aminoacid biosynthesis; cell growth regulation; chromosome aberrations; chromosome disorders; enzyme substrate complex; eukaryote; fungal genetics; gene expression; genetic manipulation; genetic mapping; genetic recombination; genetic transcription; genetic translation; histidine; molecular cloning; nucleic acid sequence; regulatory gene; structural genes; temperature sensitive mutant; transfer RNA

Our goal is to understand the regulation of gene activity in the yeast Saccharomyces cerevisiae. The studies will focus on the nucleotide sequences which regulate transcription and translation of the histidine biosynthetic enzymes. Mutations in these sequences will be obtained by in vitro and in vovo mutagenesis. The mutations derived in vivo will be cloned by recombinant DNA procedures and then sequenced; those constructed in vitro will be inserted back into the genome for study by transformation. One of the sequences responsible for regulating genes in yeast is the transposable element, TY1. This element causes loss of function by insertion into the 5' non-coding region prior to the first ATG in the gene. We will study the insertion and excision of this element genetically and biochemically. Genetically, we will study the chromosome aberrations caused by the element as well as characterize mutations which enhance or diminish its insertion and excision. We will characterize the join points between the target gene and the element as well as the specificity of the recombination event by nucleotide sequence analysis. The regulatory effort of the insertion element on the transcription capabilities of the adajcent structural gene will be studied by Northern analysis.

我们的目标是了解酵母中基因活性的调节 酿酒酵母 这些研究将集中在核苷酸序列上 其调节组氨酸生物合成的转录和翻译 内切酶 这些序列中的突变将通过体外和体内获得 诱变 体内衍生的突变将通过重组DNA克隆 程序，然后测序;那些在体外构建的将被插入回 转化到基因组中进行研究。 在酵母中负责调节基因的序列之一是 转座因子TY 1。 该元件通过插入导致功能丧失 在该基因的第一个ATG之前插入5'非编码区。 我们将研究 基因和生物化学上的插入和切除。 在遗传学上，我们将研究该元素引起的染色体畸变， 以及表征增强或减少其插入的突变， 切除 我们将描述目标基因和 元件以及核苷酸重组事件的特异性 序列分析 插入元件对细胞的调节作用 将通过以下方法研究相邻结构基因的转录能力： 北方分析。