STRUCTURE & ORGANIZATION OF YEAST MAL LOCI

拟议中的研究旨在研究分子 结构基因表达调控机制(S) 参与酵母中麦芽糖(和淀粉)的代谢。我们是 对描述顺式和反式的特征特别感兴趣 参与诱导的MAL基因座的调控成分 葡萄糖对麦芽糖和碳分解代谢的抑制作用。努力是 针对麦芽糖酶和麦芽糖的结构和功能 渗透酶启动子元件及其反式作用调节 蛋白质及其作用部位(S)。 1.完成MAL6基因座的测序，鉴定 MALR重复序列(MALRR)和测序MAL1 基因间隔区。在这些实验中，序列 完整的MAL6基因座和重复序列(MAL6RR) 与MAL6R同源的基因将完成。关于中国农业科技发展的初步研究 将启动MAL6RR的属性。 2.对MAL6-IG启动子元件的进一步研究 区域。我们将更详细地研究UASM序列 尊重它的边界，无论它的方向和距离 来自TATA序列的是重要的，因为它可能在 调控差异转录的渗透酶基因。的作用 富T序列在葡萄糖抑制中的作用有待进一步研究 并进行检查以确定特定的蛋白质是否与其结合。 3.MAL6R编码调控蛋白的研究。这个 调节蛋白将在大肠杆菌中表达并在 以便在体外制造抗体。调节蛋白 它的一些突变的衍生品将被研究 与麦芽糖酶启动子区域相互作用。 4.寻找麦芽糖酶的构成和葡萄糖抑制 不敏感的突变体。构成突变将通过以下方式产生 突变调控基因和免疫球蛋白区域。他们将会是 检查它们是否受到葡萄糖的抑制。 5.MAL1基因座重复。二倍体将被构建 这将使人们能够分析随机孢子以寻找复制的 MAL1基因座。 6.酵母糖淀粉酶的研究STA2基因的序列测定 将完成STA10基因的克隆，STA基因的定位 并研究了MATA/MATAlpha对STA基因的调控。

The proposed studies are designed to investigate the molecular mechanism(s) controlling the expression of the structural genes involved in maltose (and starch) metabolism in yeast. We are particularly interested in characterizing the cis and trans regulatory components of the MAL locus involved in induction by maltose and carbon catabolite repression by glucose. Effort is directed at the structure and function of the maltase and maltose permease promoter elements and of the trans-acting regulatory protein and its site(s) of action. 1. Complete the sequencing of the MAL6 locus, characterize the MALR repeated sequence (MALRR) and sequence the MAL1 intergenic (IG) region. In these experiments, the sequencing of the entire MAL6 locus and of a repeated sequence (MAL6RR) homologous to MAL6R will be completed. Preliminary studies on the properties of MAL6RR will be initiated. 2. Additional studies on the promoter elements in the MAL6 IG region. The UASM sequence will be studied in greater detail with respect to its boundaries, whether its orientation and distance from the TATA sequence are important and for its possible role in regulating the divergently transcribed permease gene. The role of the T-rich sequence in glucose repression will be studied further and examined to determine whether specific proteins bind to it. 3. Studies on the MAL6R encoded regulatory protein. The regulatory protein will be expressed in E. coli and synthesized in vitro in order to make antibodies to it. The regulatory protein and some of its mutationally altered derivatives will be studied for interaction with the maltase promoter region. 4. Search for maltase constitutive and glucose repression insensitive mutants. Constitutive mutants will be generated by mutagenizing the regulatory gene and the IG region. They will be checked for their repression by glucose. 5. Duplication of the MAL1 locus. Diploids will be constructed which will allow one to analyze random spores for duplications of the MAL1 locus. 6. Studies on yeast glucoamylase. Sequencing of the STA2 gene will be completed, the STA10 gene cloned, the STA genes mapped and the regulation of STA genes by MATa/MATalpha investigated.