GENOMIC ORGANIZATION OF PHOTOSYNTHETIC GENES

光合基因的基因组组织

• 批准号: 3305709
• 负责人: WILLIAM R WIDGER
• 金额: $ 11.31万
• 依托单位: UNIVERSITY OF HOUSTON
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-07-06 至 1994-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3305709
• 关键词: Cyanophyta; bacterial proteins; computer assisted sequence analysis; endonuclease; gene expression; genetic library; genetic models; genetic transcription; genome; messenger RNA; molecular cloning; northern blottings; nucleic acid hybridization; nucleic acid probes; photosynthesis; photosynthetic bacteria; polymerase chain reaction; protein structure function; pulsed field gel electrophoresis; radiotracer; regulatory gene; restriction mapping; southern blotting

The long term objective of this project is to construct a detailed restriction map of overlapping lambda clones using an existing coarse NotI restriction map of the cyanobacterium, Agmenellum quadruplicatum PCC 7002. This map will allow us to understand how the Agmenellum genome controls photosynthesis. Tbe contig mapping experiments takes advantage of a technique used to group lambda clones from a overlapping clone library into ensembles of clones. Each ensemble belongs to a unique large pulsed field restriction fragment. Tbe advantages to grouping clones into ensembles are: a) a limited number of clones are needed to construct a contig, b) contig assembly proceeds along the physical restriction map of an organism, c) and ensembles of clones derived from large overlapping pulsed field restriction fragments can be used to create physical restriction maps. Once the detailed physical map is constructed, a genetic map can be made using hybridization probes of existing cyanobacterial clones. The organization of photosynthetic genes in relationship to other important genes of biochemical pathways such as nitrogen metabolism (glnA), amino acid biosynthesis, carbohydrate synthesis (Calvin cycl), protein synthesis (rrnA, rrnB, rps genes) and RNA and DNA metabolism (rpo genes, recA, and AquI methylase) will be compared using genomic transcript mapping. Light controls the growth and replication of this bacterium, in darkness it will down regulate photosynthetic pathways by diminishing transcripts of these genes. Upon illumination these gene transcripts become overly abundant reaching levels several fold greater than exists in steady state fight. The differential response of transcripts to light will be mapped on the genome by screening a separated and ordered clone library from labelled transcripts isolated in the dark and in the Eght. The techniques used here in should be applicable to map many prokaryotic genomes including those of pathogenic organisms whose detailed genomic organization or expression of certain genes are involved in a toxigenic or pathogenic phenotype.

这个项目的长期目标是建造一个详细的 使用现有粗集的重叠的lambda克隆的限制图 四倍体蓝藻的NOTI限制性内切酶图谱 PCC 7002。这张地图将让我们了解Agmenellum是如何 基因组控制光合作用。重叠群作图实验所需时间 用于将重叠的lambda克隆分组的技术的优点 将文库克隆成克隆的集合。每个乐团都属于一个 唯一的大型脉冲场限制片段。具有以下优势： 将克隆分组为合奏：a)有限数量的克隆是 构建重叠群所需的，b)重叠群组装沿着 生物体的物理限制图，c)和克隆的集合 从大的重叠脉冲场限制片段得到的可以是 用于创建物理限制图。一旦详细的体检 构建了遗传图谱，就可以用杂交探针绘制遗传图谱 现有的蓝藻克隆。光合作用的组织 生化途径中与其他重要基因相关的基因 如氮代谢(GlnA)、氨基酸生物合成、 碳水化合物合成(卡尔文周期)、蛋白质合成(rRNA、rrnB、RPS 基因)和RNA和DNA代谢(rpo基因、recA和Aqui甲基酶) 将使用基因组转录图谱进行比较。灯光控制着 这种细菌的生长和复制，在黑暗中它会下降 通过减少这些基因的转录来调节光合作用途径 基因。在光照下，这些基因转录物变得过于丰富。 达到比稳态战斗多几倍的等级。 转录本对光的不同反应将被绘制在 从标记的克隆文库中筛选分离有序的克隆文库 在黑暗和光明中被隔离的文字记录。 本文使用的技术应该适用于对许多原核生物进行作图 基因组包括其详细基因组的病原生物体的基因组 某些基因的组织或表达与产毒有关 或致病表型。