GENETIC ANALYSIS OF BACTERIAL PHOTOSYNTHESIS

细菌光合作用的遗传分析

• 批准号: 3279883
• 负责人: SAMUEL KAPLAN
• 金额: $ 20.73万
• 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/3279883
• 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; 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.

利用兼性光养细菌，球形红球藻， 我们正试图在分子水平上了解基因控制 诱导型光合膜系统（ICM）。 的遗传基础 ICM表达式扩展到：1.主要细菌叶绿素蛋白 光谱复合物、细胞色素、ATP酶和其他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.磷脂生物合成酶与 磷脂的运动，和4.监管制度导致 细胞对氧分压和光照变化的适应 强度，以及那些有序组装的机制， 以及ICM的结构-功能相互关系。 是 我建议我们识别、克隆和描述这些基因区域， 与ICM结构和监管相关。 该研究包括，结构 通过限制性内切酶分析， 测序以及基因表达的体外和体内表征。 我们建议制定选择、分离和 在体内和体外表征突变， 与ICM结构和功能相关， 脱氧寡核苷酸和转座子诱导的诱变。 我们打算 衍生遗传系统来监测这些克隆区域的表达， 同源和异源宿主，以便更好地了解 基因控制机制。 这些研究将直接影响到更广泛的 调节信号的性质及其在 异源宿主系统 最后，我们打算发展基因和 假遗传系统的运动，互补，表达和 对R. sphaeroides 我们将试图 开发接合和转导系统、连接酶、质粒、启动子 克隆载体等等，采用经典的原核遗传技术 以及重组DNA技术。 了解功能和 光合作用的结构影响是充分利用 营养，因此，人类的健康。 平等 重要性，结构-功能相互关系 膜相关活动对健康至关重要，因为 与健康相关的膜现象，例如膜受体， 线粒体功能、排泄、脂蛋白结构和功能等。