GENE EXPRESSION AND CONTROL OF INTERMEDIATE METABOLISM

我们的目标是了解协调一组 基因。 我们已经选择了研究的基因编码的酶， 糖酵解途径 这条通道包含许多井 特征代谢步骤，其主要目的是产生 碳水化合物的能量。 此外，几种中间体具有 替代代谢命运，最显著的是脂肪酸生物合成。 我们 建议继续我们对表达和结构的描述 甘油-3-磷酸脱氢酶 甘油醛-3-磷酸脱氢酶 异构酶（TPI）和β-羟酸脱氢酶（HAD）。 前三 酶连接了一系列反应之间的上部 糖酵解、脂肪酸生物合成和糖酵解的下部。 的 每一种基因都已分离出来，他们的研究将继续下去。 我们 计划一系列实验，以确定是否酶的基因上的酶， 糖酵解途径作为一个集合被控制， 彼此独立地。 转录本和酶含量将 在不同的细胞类型中测量，在饮食被改变的动物中 碳水化合物、氨基酸或脂质补充剂，以及在不同物种中 生活在不同栖息地的果蝇。 我们还计划 以分离编码其他糖酵解酶的额外基因。 我们 策略将是纯化蛋白质，制备抗血清并筛选 将果蝇cDNA的重组DNA文库克隆到表达载体中， 矢量Lambdagt 11。 这种方法已成功地用于分离 我们正在研究的基因。 每种基因的克隆和抗体 蛋白质，我们将开始研究，以确定DNA序列是网站 的调控，并确定转录因子，影响 每个基因和整个基因组的表达。

Our goals are to understand the mechanism which operate to coordinate a set of genes. We have selected for study the genes which encode the enzymes of the glycolytic pathway. This pathway contains a number of well characterized metabolic steps whose principal end is the generation of energy from carbohydrates. In addition several intermediates have alternative metabolic fates most notably fatty acid biosynthesis. We are proposing to continue our characterization of the expression and structure of the genes which code for the enzymes, glycerol-3-phosphate dehydrogenase (GPDH); glyceraldehyde-3-phosphate dehydrogenase (GAPDH); triose phosphate isomerase (TPI) and Beta-hydroxyacid dehydrogenase (HAD). The first three enzymes interconnect a series of reactions between the upper part of glycolysis, fatty acid biosynthesis and the lower part of glycolysis. The genes for each have been isolated and their study will continue. We are planning a set of experiments to ascertain if the genes for enzymes on the glycolytic pathways are controlled as a set or whether they are controlled independently of one another. Transcripts and enzyme content will be measured in different cell types, in animals whose diet has been altered by carbohydrate, amino acid or lipid supplementation, and in different species of Drosophila that live in very different habitats. We are also planning to isolate the additional genes which encode other glycolytic enzymes. Our strategy will be to purify the protein, prepare an antiserum and screen recombinant DNA libraries of Drosophila cDNA cloned into the expression vector Lambdagt11. This approach has been used successfully to isolate the genes we are now studying. With clones of each gene and antibody for each protein we would begin studies to identify DNA sequences which are the site of regulation and to identify transcriptional factors which affect the expression of each of the genes and the entire set.