STUDIES ON FLAGELLAR MORPHOGENESIS

鞭毛形态发生的研究

• 批准号: 3294174
• 负责人: BESSIE P HUANG
• 金额: $ 6.94万
• 依托单位: SCRIPPS CLINIC AND RESEARCH FOUNDATION
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-07-01 至 1987-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3294174
• 关键词: Chlorophyta; affinity chromatography; antibody formation; electron microscopy; eukaryote; flagellum; gel electrophoresis; gene interaction; histogenesis; hybridomas; immunoelectron microscopy; immunofluorescence technique; immunoprecipitation; macromolecule; microscopy; monoclonal antibody; plant genetics; radiotracer; regulatory gene; structural genes; temperature sensitive mutant

The major objectives of this proposal are to study the genetic, biosynthetic, and macromolecular assembly processes involved in the morphogenesis of the eukaryotic flagellum. The proposed studies will be carried out in the biflagel-late alga, Chlamydomonas reinhardtii, because of its proven advantages as a model system for molecular analysis of flagellar structure and assembly. Given the structural and biochemical complexity of the flagellum, molecular dissection of its biogenesis requires that highly specific and sensitive probes of the system area available. This research project is directed toward generating and utilizing such immunological and genetical probes. A library of hybridomas secreting monoclonal antibodies have been generated already in our laboratory, and additional ones will be isolated. These monoclonal antibodies will be specifically used to follow different flagellar proteins from their time of synthesis to assembly into structure. These studies will utilize the rapid and synchronous regeneration of flagella which occurs in Chlamydomonas following experimental deflagellation. Concurrent with these studies, we will pursue a molecular and genetic characterization of a series of non-conditional and thermosensitive mutants of Chlamydomonas with specific defects for flagellar biogenesis. Through our analysis of these mutants, already at hand, we anticipate that structural and regulatory genes which play critical functions in flagellar morphogenesis will be identified.

该提案的主要目标是研究遗传、 生物合成和大分子组装过程涉及 真核生物鞭毛的形态发生。 拟议的研究将是 在双鞭毛藻莱茵衣藻中进行，因为 作为分子分析模型系统，其已被证实的优势 鞭毛结构和组装。 鉴于结构和生化 鞭毛的复杂性，其生物起源的分子解剖 要求系统区域具有高度特异性和灵敏性的探针 可用的。 该研究项目旨在产生和 利用此类免疫学和遗传探针。 杂交瘤文库 分泌单克隆抗体已经在我们的 实验室，其他人将被隔离。 这些单克隆 抗体将专门用于追踪不同的鞭毛蛋白 从合成到组装成结构的时间。 这些研究 将利用鞭毛的快速同步再生 发生在衣藻中实验性爆鞭之后。 并发 通过这些研究，我们将寻求分子和遗传特征 衣藻系列非条件热敏突变体的研究 具有鞭毛生物发生的特定缺陷。 通过我们的分析 这些突变体已经存在，我们预计结构和 在鞭毛形态发生中起关键作用的调控基因 将被识别。