PHOTOSYNTHESIS, MUTANTS, AND CLONING IN CYANOBACTERIA

蓝藻的光合作用、突变体和克隆

• 批准号: 3270726
• 负责人: LOUIS A SHERMAN
• 金额: $ 11.72万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-05-01 至 1989-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3270726
• 关键词: Escherichia coli; bacterial genetics; chlorophyll; conformation; electron microscopy; fluorescence; gel electrophoresis; genetic manipulation; genetic mapping; genetic recombination; membrane structure; molecular cloning; photochemistry; photosynthesis; photosynthetic bacteria; radiotracer; temperature sensitive mutant

The purpose of this proposal is to understand the relationship between membrane architecture and function in photosynthetic membranes. Our approach will be via discrete, yet correlative, techniques using the unicellular, transformable cyanobacterium Anacystis nidulans R2. The emphasis will be on Photosystem II and the oxygen-evolving apparatus. Our strategy will consist of seven major experimental techniques: (1) the isolation and characterization of an oxygen-evolving complex that contains Photosystem II and phycobilisomes; (2) site-specific labeling and cross-linking of proteins to determine their position in the membrane; (3) reconstitution of photosynthetic electron transport from H2O to NADP+ in liposomes using isolated complexes; (4) isolation of photosynthetic mutants and herbicide-resistant mutants. In addition to temperature-sensitive mutants, strains will be isolated that must be maintained by heterotrophic growth; (5) complementation analysis with cloned genes; (6) cloned photosynthesis genes will be used for biochemical and genetic experiments; and (7) fluorescence analysis of chlorophyllprotein complexes in normal and developing cells. This project is a combined genetic and biochemical study of membrane structure and function and is thus of great benefit to membrane biology in general. The specific information that should be obtained from this proposal includes: (1) the protein composition of the major membrane complexes, such as the O2-evolving apparatus and chlorophyll-proteins; (2) the precise location of proteins in the membrane; (3) the effects of mutation on membrane structure and function; (4) the effect of adding cloned photosynthesis genes to wild-type and mutant cells; (5) the identity of genes coding for photosynthetic proteins; (6) the identity of the chlorophyll components that give rise to the 77 K fluorescence emission peaks; and (7) the mechanism of membrane assembly in iron-deficient cells after iron addition and in heterotrophic cells after light induction. The combination of biochemistry and genetics makes A. nidulans a very valuable organism for the study of bioenergetics and membrane structure.

这项建议的目的是了解 光合膜中的膜结构和功能。我们的 方法将通过离散的、但相关的技术使用 单细胞可转化蓝藻Anacystis nidulans R2。这个 重点将放在光系统II和放氧装置上。我们的 战略将由七项主要实验技术组成：(1) 一种含氧化合物的放氧复合体的分离和鉴定 光系统II和藻胆体；(2)位点特异性标记和 蛋白质的交联以确定它们在膜中的位置；(3) InH2O向NaDP+光合作用电子传递的重建 利用分离的复合体制备脂质体；(4)光合作用突变体的分离 以及抗除草剂突变体。除了对温度敏感之外 突变株，菌株将被分离出来，必须通过异养方式维持 生长；(5)与克隆基因的互补分析；(6)克隆 光合作用基因将用于生化和遗传实验； 以及(7)正常和正常的叶绿素蛋白复合体的荧光分析。 发育中的细胞。 本项目是膜的遗传学和生物化学相结合的研究。 结构和功能，从而对膜生物学有很大的帮助 将军。应从此获取的特定信息 建议包括：(1)主膜的蛋白质组成 放氧器和叶绿素蛋白等复合体；(2) 蛋白质在膜中的精确定位；(3) 突变对膜结构和功能的影响；(4)添加 将光合作用基因克隆到野生型和突变型细胞；(5)同源性 编码光合作用蛋白的基因；(6) 引起77K荧光发射的叶绿素组分 (7)缺铁细胞膜组装机制。 加铁后异养细胞和光诱导后异养细胞。这个 生物化学和遗传学的结合使根结线虫成为一种非常有价值的 用于研究生物能量学和膜结构的有机体。