PROLIFERATION-SPECIFIC GENES ANALYSIS IN DICTYOSTELIUM

盘基菌增殖特异性基因分析

• 批准号: 3186464
• 负责人: WILL J KOPACHIK
• 金额: $ 6.34万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-06-01 至 1990-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3186464
• 关键词: Dictyostelium; autoradiography; cell cycle; cell differentiation; cell growth regulation; complementary DNA; cyclic AMP; developmental genetics; endonuclease; fungal genetics; gene expression; genetic regulation; growth /development; molecular cloning; neoplastic growth; nucleic acid hybridization; nucleic acid probes; nucleic acid sequence; temperature sensitive mutant

The cellular slime mold, Dictyostelium discoideum, is an ideal organism for research on the control of cell proliferation. In its life cycle the vegetative amoebae first proliferate and then become quiescent as they differentiate in response to starvation. Genetic and molecular analysis of the transition revealed three classes of genes: "vital" genes expressed at all stages, proliferation genes selectively expressed only during cell proliferation and differentiation genes expressed only during development. The putative proliferation genes could include a novel set of genes essential for DNA synthesis, mitosis, cytokinesis or other aspects of cell proliferation. In spite of considerable effort in other systems relatively few cell division cycle genes have yet been found. The major objectives in the research include using cDNA probes of the three classes comparatively to determine specific mechanisms of proliferation gene expression. Filter hybridization methods will be used to identify the most likely candidates by their regulation and conservation in evolution. Some of the genes have homologies to mammalian sequences. Additional homologies to known cdc genes will be sought by hybridization studies and sequence analysis. The turn off of the genes is under genetic control. The mutations may be in regulatory factors which control cdc gene expression. Thermosensitive conditional mutants will be isolated by a novel selection protocol. A current hypothesis that cAMP represses these gene's expression will be assessed by RNA blot analysis of cAMP chemosensory mutants. The final result will be an identification of known and novel proliferation genes and an identification of genes and physiological events which control their expression. Although mammalian cells share some of the same genes studied here, they lack the experimental advantages of this system. Therefore, knowledge about the functions and regulation of these genes may aid in understanding developmental diseases and neoplastic growth.

细胞粘液模具，dictyostelium discoideum，是一种理想的生物体 研究细胞增殖的控制。 在其生命周期中 营养变形虫首先扩散，然后在它们时变为静止 响应饥饿而区分。 遗传和分子分析 过渡揭示了三类基因：表达的“重要”基因 所有阶段，仅在细胞中选择性表达的增殖基因 增殖和分化基因仅在发育过程中表达。 假定的增殖基因可能包括一组新的基因 对于DNA合成，有丝分裂，细胞因子或细胞的其他方面必不可少的 增殖。 尽管在其他系统中付出了巨大的努力 几乎没有发现细胞分裂周期基因。 主要目标 研究包括使用三类的cDNA探针 确定增殖基因表达的特定机制。 筛选 杂交方法将用于识别最可能的候选人 通过他们在进化中的监管和保护。 一些基因具有 哺乳动物序列的同源性。 已知CDC的其他同源 杂交研究和序列分析将寻求基因。 这 关闭基因在遗传控制下。 突变可能在 控制CDC基因表达的调节因子。 热敏性 有条件的突变体将通过新颖的选择方案隔离。 一个 当前cAMP抑制这些基因表达的假设将是 通过CAMP化学感突变体的RNA印迹分析评估。 决赛 结果将是已知和新颖的增殖基因的鉴定以及 控制其基因和生理事件的识别 表达。 尽管哺乳动物细胞共享研究的一些相同基因 在这里，他们缺乏该系统的实验优势。 所以， 有关这些基因功能和调节的知识可能有助于 了解发育疾病和肿瘤生长。