REGULATION OF HISTIDINE BIOSYNTHESIS IN YEAST

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 它们调节组氨酸生物合成的转录和翻译 酵素。这些序列的突变将通过体外和体内获得 诱变。体内产生的突变将通过重组DNA进行克隆 程序，然后测序；那些在体外构建的将被插入回 通过转化进入基因组进行研究。 在酵母中负责调节基因的序列之一是 转座元件，Ty1。此元素因插入而导致功能丧失 插入到基因中第一个ATG之前的5‘非编码区。我们会研究 从基因和生物化学的角度插入和切除这种元素。 在遗传学方面，我们将研究这种元素引起的染色体异常 以及表征增强或减少其插入的突变以及 切除。我们将描述目标基因和基因之间的连接点 元素以及核苷酸重组事件的特异性 序列分析。插入元件对 将通过以下方法研究邻近结构基因的转录能力 北方分析。

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.