BIOCHEMISTRY AND GENETICS OF MEMBRANE FUNCTIONS

我们以前已经证明，E.大肠杆菌与 与氨基酸的主动转运和某些简单的 - 是的 由于其在细胞过程中的重要性， 这种能量耦合因子（ECF）的遗传特性将被 追求。 ECF与β-胱硫醚酶的结构关系将 通过DNA测序在核苷酸水平上进行研究和阐明。 同时携带ecf和metC（胱硫醚酶）基因的克隆已被分离出来 并将用于这些研究。 提出了实验来解决 渗透冲击敏感输送的能量学问题 系统. 将尝试确定直接能量供体， 这些运输系统通过找到第三个反应， 磷酸盐和研究乙酰磷酸盐的命运。 谷氨酰胺 运输系统E. coli中的质粒pBR 322。 利用这一点，遗传结构以及分子组织的 将确定并阐明该传输系统。 重建 谷氨酰胺转运将在原生质球中进行 转运突变体作为体外互补试验， 所涉及的运输组件的功能作用。 生理 谷氨酰胺转运缺陷对谷氨酸催化的影响 将在最初阶段进行遗传学研究，然后进行生物化学研究。 研究找出这种效应的生物化学基础是什么。 主要 这项研究的目的是扩大我们的知识， 能量转换机制的分子和遗传组织 以及代谢能与活性物质偶联的分子机制 运输

We have shown previously that the ecf gene of E. coli is intimately associated with the active transport of amino acids and certain simple saccharides. Because of its importance in cellular processes, biochemical and genetic characterization of this energy coupling factor (ECF) will be pursued. The structural relationship of the ECF to beta-cystathionase will be investigated and elucidated at the nucleotide level, by DNA sequencing. Clones carrying both ecf and metC (cystathionase) gens have been isolated and will be used for these studies. Experiments are proposed to tackle the problem of the energetics of the osmotic shock-sensitive transport systems. Attempts will be made to identify the direct energy donor for these transport systems by finding the third reaction that makes acetyl phosphate and by investigating the fate of acetyl phosphate. The glutamine transport system of E. coli has been cloned using the plasmid pBR322. Using this, the genetic structure as well as the molecular organization of this transport system will be determined and elucidated. Reconstitution of glutamine transport will spheroplasts will be carried out with glutamine transport mutants as an in vitro complementation test to sort out the functional role of the transport components involved. The physiological effects of the glutamine transport defect on the catabolism of glutamate will be purused genetically in the initial phase, followed by biochemical study to find out what is the biochemical basis of the effect. The main objective of this research proposal is to broaden our knowledge concerning the molecular and genetic organization of the energy transduction machinery and the molecular mechanisms of the coupling of metabolic energy to active transport.