GENETIC ANALYSIS OF BACTERIAL PHOTOSYNTHESIS

细菌光合作用的遗传分析

• 批准号: 3279880
• 负责人: SAMUEL KAPLAN
• 金额: $ 21.98万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-08-01 至 1991-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3279880
• 关键词: Escherichia coli; Rhodopseudomonas; bacterial DNA; bacterial genetics; electron transport; gene expression; genetic manipulation; genetic regulation; laboratory rabbit; membrane activity; membrane structure; microorganism culture; molecular cloning; mutant; nucleic acid probes; nucleic acid sequence; operon; photosynthesis; photosynthetic bacteria; plasmids; pseudogenes; structural genes

Using the facultative phototrophic bacterium, Rhodopseudomonas sphaeroides, we are attempting to understand, at the molecular level, gene control of the inducible photosynthetic membrane system (ICM). The genetic basis for ICM expression extends to the: 1. Major bacteriochlorophyll-protein spectral complexes, cytochromes, ATPase and other ICM-associated enzyme activities and structural proteins, 2. The pathways for carotenoid and bacteriochlorophyll synthesis, 3. Phospholipid biosynthetic enzymes and the movement of phospholipids, and 4. The regulatory systems resulting in cellular adaptation to changes in oxygen partial pressure and light intensity, and those mechanisms involved in assembly in an orderly manner, as well as the structure-function interrelationships of the ICM. It is proposed that we identify, clone and characterize those genetic regions relevant to ICM structure and regulation. This study includes, structure of the DNA region, through restriction endonuclease analysis, DNA sequencing and in vitro and in vivo characterization of gene expression. We propose to develop procedures for the selection, isolation and characterization, both in vivo and in vitro, of mutations which are relevant to ICM structure and function by employing synthetic deoxyoligonucleotide and transposon induced mutagenesis. We intend to derive genetic systems to monitor the expression of these cloned regions in both homologous and heterologous hosts in order to better understand the mechanisms of gene control. Such studies will bear directly on the broader question of the nature of the regulatory signals and their function in heterologous host systems. Finally, we intend to develop genetic and pseudogenetic systems for the movement, complementation, expression and facile handling of genetic regions in R. sphaeroides. We shall attempt to develop conjugation and transduction systems, cosmids, plasmids, promoter cloning vehicles, etc., employing classical procaryotic genetic techniques as well as recombinant DNA technologies. Understanding the functional and structural implications of photosynthesis is fundamental to the adequate nutrition and, therefore, health of human populations. Of equal importance, the structure-function interrelationships of membrane-associated activities is of critical health importance because of the nature of health related membrane phenomena, e.g. membrane receptors, mitochondrial function, excretion, lipoprotein structure and function, etc.

利用兼性光营养细菌球形红假单胞菌， 我们正试图在分子水平上理解基因控制 诱导光合膜系统(ICM)。人类的遗传基础 ICM的表达延伸到：1.主要的细菌叶绿素蛋白 光谱复合体、细胞色素、ATPase和其他ICM相关酶 活性和结构蛋白，2.类胡萝卜素和 细菌-叶绿素合成，3.磷脂生物合成酶和 磷脂的运动，以及4.导致 细胞对氧分压和光照变化的适应 强度，以及那些有序地参与组装的机构， 以及ICM的结构-功能相互关系。它是 建议我们识别、克隆和表征这些基因区域 与ICM结构和法规相关。这项研究包括，结构 通过限制性内切酶分析，DNA区域的 基因表达的测序和体外和体内表征。 我们建议制定程序，以选择、隔离和 在体内和体外，突变的特征是 与ICM结构和功能相关的应用 脱氧寡核苷酸和转座子诱导的突变。我们打算 衍生遗传系统以监测这些克隆区域在 同源和异源寄主，以便更好地了解 基因控制的机制。这样的研究将直接影响到更广泛的 关于调节信号的性质及其在 异种寄主系统。最后，我们打算开发基因和 运动、互补、表达和表达的伪遗传系统 球状乳杆菌遗传区的简易处理。我们将尝试 发展接合和转导系统、粘粒、质粒、启动子 使用经典原核基因技术的克隆工具等 以及重组DNA技术。了解功能和 光合作用的结构含义是充分利用 营养，因此，人类人口的健康。平等的 重要性，结构与功能的相互关系 膜相关活动对健康至关重要，因为 与健康有关的膜现象的性质，例如膜受体， 线粒体功能、排泄、脂蛋白结构和功能等。