GENE EXPRESSION AND CONTROL OF INTERMEDIATE METABOLISM

我们的目标是了解协调一套设备的运行机制 基因。我们选择了编码人乳酸酶的基因进行研究。 糖酵解途径。这条小路上有许多井 以代谢步骤为特征，其主要目的是产生 来自碳水化合物的能量。此外，还有几个中间体 另类代谢命运，最显著的是脂肪酸生物合成。我们是 建议继续我们对表达和结构的描述 在编码这些酶的基因中，甘油-3-磷酸脱氢酶 甘油醛-3-磷酸脱氢酶；磷酸三糖 异构酶(TPI)和β-羟基酸脱氢酶(HAD)。前三名 酶使上半部分之间的一系列反应相互连接 糖酵解、脂肪酸生物合成和糖酵解的下部。这个 每个人的基因都已经分离出来，他们的研究将继续进行。我们是 计划进行一系列实验，以确定细胞上的酶基因 糖酵解途径被控制为一组，或者它们是否被控制 彼此独立。成绩单和酶含量将是 在饮食被改变的动物中，以不同的细胞类型进行测量 碳水化合物、氨基酸或脂肪补充剂，以及不同种类的 生活在非常不同的栖息地的果蝇。我们也在计划 分离编码其他糖酵解酶的额外基因。我们的 策略将是提纯蛋白质，制备抗血清并筛选 果蝇重组DNA文库的克隆及其在大肠杆菌中的表达 载体Lambdagt11.这种方法已成功地用于分离 我们现在正在研究基因。每个基因和抗体都有克隆 蛋白质，我们将开始研究，以确定哪些DNA序列是 并确定影响转录因子的转录因子。 每一个基因和整个集合的表达。

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.