SYNECHOCYSTIS

集胞藻属

• 批准号: 7721175
• 负责人: ROBERT W ROBERSON
• 金额: $ 1.62万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-12-01 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7721175
• 关键词: Address; Animal Model; Biochemical Genetics; Biogenesis; Caliber; Cell membrane; Cells; Characteristics; Computer Retrieval of Information on Scientific Projects Database; Cyanobacterium; Cytoplasmic Structures; Data; Freeze Fracturing; Freeze Substitution; Freezing; Funding; Genome; Grant; Hand; Image; Institution; Investigation; Light; Membrane; Methods; Microtomy; Molecular Genetics; Photosynthesis; Physiological; Plant Resins; Positioning Attribute; Preparation; Research; Research Personnel; Resources; Sampling; Scanning Electron Microscopy; Site; Solar Energy; Source; Synechocystis; System; Thick; Thylakoid Membranes; Thylakoids; United States National Institutes of Health; Work; bioimaging; cell type; electron tomography; improved; membrane biogenesis; mutant; pressure; reconstruction; tomography

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The unicellular cyanobacterium Synechocystis sp. PCC 6803 is a model organism that has highly desirable molecular genetic (e.g., the genome is sequenced (Kaneko et al. 1996), easily transformed), physiological (e.g., easily grown in pure culture), and morphological (e.g., cells are ~1.5 um in diameter and thus suitable for quantitative three-dimensional (3D) ultrastructural analysis) characteristics. These characteristics make Synechocystis an ideal experimental system to address our long term questions regarding photosynthesis and the biogenesis and organization of thylakoid membranes, the sites of solar energy capture and energy transduction. There remain key unanswered questions regarding the biogenesis and 3D organization of the thylakoid membrane network in Synechocystis cells. In past work, we combined cryo-preparation methods for ultrastructural investigations, including high-pressure freezing and freeze substitution, with electron tomography of resin embedded semi-thick (250 nm) samples, serial thin section (60 nm) reconstruction analysis, and freeze fracture field-emission cryo-scanning electron microscopy to better understand cytoplasmic details in wild type (van de Meen et al. 2006) and mutant (Mohamed et al. 2005) strains. We hope to make use of cryo-EM, including tomography, at the National Center Macromolecular Imaging to image whole, wild type cells in their frozen hydrated states in order to augment our ongoing research. We believe that, if successful, the advanced bioimaging methods of cryo-EM and post-acquisition analysis will help greatly in clearing up questions regarding thylakoid membrane organization and its associations with thylakoid centers (i.e., cytoplasmic structures that support the 3D organization of the membrane network) and improve our understanding of how the thylakoid membrane system interacts with the plasma membrane. With this data in hand, along with our genetic and biochemical approaches, we will be in a much stronger position to identify sites of membrane biogenesis and understand how the 3D order of these membranes is maintained in this model organism. Results from this work could be used in shedding light in a broader sense on questions related to prokaryotic membrane biogenesis.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 单细胞蓝细菌集胞藻属PCC 6803是具有高度期望的分子遗传学（例如，基因组测序（Kaneko等1996），容易转化），生理学（例如，易于在纯培养物中生长），和形态学（例如，细胞直径约为1.5 μ m，因此适合于定量三维（3D）超微结构分析）特征。这些特性使集胞藻成为一个理想的实验系统，以解决我们长期的问题，光合作用和类囊体膜的生物起源和组织，太阳能捕获和能量转导的网站。 关于集胞藻细胞中类囊体膜网络的生物起源和3D组织，仍然存在关键的未回答的问题。在过去的工作中，我们将用于超微结构研究的冷冻制备方法，包括高压冷冻和冷冻置换，与树脂包埋的半厚（250 nm）样品的电子断层扫描，连续薄切片（60 nm）重建分析，和冷冻断裂场发射冷冻扫描电子显微镜，以更好地了解野生型细胞质的细节（货车de Meen et al. 2006）和突变体（Mohamed et al. 2005）菌株。我们希望在国家高分子成像中心利用cryo-EM，包括断层扫描，对处于冷冻水合状态的整个野生型细胞进行成像，以增强我们正在进行的研究。我们相信，如果成功的话，冷冻电镜和采集后分析的先进生物成像方法将大大有助于澄清有关类囊体膜组织及其与类囊体中心的联系（即，支持膜网络的3D组织的细胞质结构），并提高我们对类囊体膜系统如何与质膜相互作用的理解。有了这些数据，沿着我们的遗传和生物化学方法，我们将处于更有利的地位，以确定膜生物发生的网站，并了解这些膜的3D秩序是如何在这个模型生物体保持。这项工作的结果可以用于在更广泛的意义上阐明与原核生物膜生物发生有关的问题。