CONTROL OF GENE ACTIVITY AND ORGANELLE BIOGENESIS

基因活性和细胞器生物发生的控制

• 批准号: 3274429
• 负责人: Martin A Gorovsky
• 金额: $ 18.05万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-07-01 至 1988-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3274429
• 关键词: Tetrahymena; autoradiography; cell differentiation; cell free system; chromatin; cilium; clone cells; developmental genetics; gel electrophoresis; gene expression; genetic manipulation; genetic mapping; genetic recombination; genetic transcription; genetic translation; immunochemistry; messenger RNA; microorganism genetics; molecular cloning; mutant; nucleic acid sequence; organelles; radiotracer; regeneration

The long-term goal of this project is to determine how a signal originating at the cell periphery triggers a series of events culminating in the synthesis and assembly of an intracellular organelle. Specifically, we are studying the mechanisms by which deciliation of the ciliated protozoan Tetrahymena thermophila initiates the orderly expression of specific genes required for reformation of cilia. Improved methods have been developed for obtaining viable deciliation of Tetrahymena which induces a rapid (approximately l0-fold) induction of protein synthesis and a specific, relative (approximately l0-fold) co-ordinate induction of alpha and beta tubulins. Translational and transcriptional mechanisms underlying this general induction of protein synthesis and specific induction of tubulin are being examined. Polysomal polyA ion RNA isolated from regenerating cells 80-l00 min. after deciliation (at which time tubulin represents approximately l0% of total protein synthesis) is being utilized as a source of cDNA from which tubulin gene sequences will be prepared by recombinant DNA techniques. These cloned DNA sequences will then be utilized to study the organization of tubulin genes and their expression during regeneration.

该项目的长期目标是确定信号如何在 细胞外围触发了一系列事件，最终导致合成和 细胞内细胞器的组装。具体地说，我们正在研究 纤毛原生动物四膜虫的降解机制 嗜热菌启动特定基因的有序表达 纤毛的改造。已经开发出改进的方法来获得可行的 四膜虫分离，导致快速(约10倍) 诱导蛋白质合成和一种特定的、相对的(约10倍) 协调诱导α和β微管蛋白。翻译和 这种蛋白质的普遍诱导背后的转录机制 微管蛋白的合成和特异性诱导正在研究中。多体多角体 从再生细胞中分离的离子RNA为80-100分钟。在分治之后(在什么时候 微管蛋白约占总蛋白质合成的10%) 用作制备微管蛋白基因序列的cDNA源 通过重组DNA技术。然后将利用这些克隆的DNA序列 研究微管蛋白基因的组织和表达 再生。