GLOBAL GENE EXPRESSION IN E COLI--NITROGEN METABOLISM

我们将研究E.大肠杆菌中的基因表达。特别是，我们将研究氮调节途径及其通过常见中间体谷氨酸与渗透调节途径的联系。我们将创建E.大肠杆菌基因的荧光标记方法。colicDNA。作为第一个练习，我们将使用微阵列来确定哪些基因在大肠杆菌中受氮调节C（NtrC）蛋白的调节控制。杆菌然后，我们将研究在氮限制条件下所有基因的表达水平，以确定氮限制如何传播的主要谷氨酰胺限制下游，以减缓大肠杆菌的生长速度。杆菌最后，我们将研究谷氨酸和K+在细胞内谷氨酸调节中的作用以及谷氨酸在谷氨酸调节和生物合成中的双重作用。我们开发的细菌基因表达监测系统将有助于细菌的全局表达分析。

We will investigate regulatory pathways in E. coli by monitoring global gene expression on microarrays. In particular, we will study nitrogen regulatory pathways and their connection to osmoregulatory pathways through the common intermediate glutamate. We will create microarrays of E. coli genes and develop methods for fluorescent labeling of E. coli cDNAs. As a first exercise, we will use microarrays to determine which genes are under regulatory control of the nitrogen regulatory C (NtrC ) protein in E. coli. Then we will investigate expression levels of all genes under nitrogen limiting conditions to determine how nitrogen limitation propagates downstream of a primary glutamine limitation to slow the growth rate of E. coli. Finally, we will investigate the roles of glutamate and K+ in cellular osmoregulation and the dual role of glutamate in osmoregulation and biosynthesis. Our development of a system for gene expression monitoring in bacteria will facilitate global expression analyses in bacteria.