GENETIC STUDIES OF NON-ERYTHROID SPECTRIN

非红细胞血影蛋白的遗传学研究

• 批准号: 3296790
• 负责人: DANIEL none BRANTON
• 金额: $ 24.1万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-06-01 至 1993-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3296790
• 关键词: Drosophilidae; binding proteins; cell differentiation; chemical binding; complementary DNA; developmental genetics; electron microscopy; enzyme linked immunosorbent assay; gel electrophoresis; gene complementation; gene expression; genetic library; genetic mapping; laboratory mouse; laboratory rabbit; membrane activity; membrane proteins; molecular cloning; monoclonal antibody; morphology; mutagen testing; nucleic acid probes; nucleic acid sequence; spectrin; temperature sensitive mutant; tissue /cell culture

Our long term goal is to define the characteristic cellular and developmental role or roles of spectrin and understand how spectrin molecules contribute to specific cellular functions. We believe our long term goal can be attained by using a model system, Drosophila melanogaster, that can be investigated at both the cellular and the tissue level using a combination of biochemical, structural and genetic approaches. Antibodies will be used to study the distribution and expression of spectrin both in cell culture and during development and to identify DNA sequences that express Drosophila spectrin. Clones of these DNA sequences will be used to map the physical location of the spectrin coding sequences on the genetic map of Drosophila and classical genetic analysis and in vitro mutant production will then be carried out. The mutants will be used to investigate spectrin function in vivo and to determine the effects of the complete absence of functional spectrin. The clones will also be used together with the technology of anti-sense inhibition of expression to inhibit spectrin expression in cultured cells so that we may probe the effects of the absence of functional spectrin or the consequences of mutant spectrin expression in cultured cells. Spectrin is present in nearly all human cells but the health consequences of defective spectrin expression have only been explored in erythrocytes. The proposed experiments should provide an in-depth understanding of spectrin's role in non- erythroid cells.

我们的长期目标是确定特征细胞和 血影蛋白的发展作用或作用，并了解如何 血影蛋白分子有助于特定的细胞功能。 我们 我相信我们的长期目标可以通过使用一个模型来实现， 系统，果蝇，可以在这两个调查 在细胞和组织水平上， 生物化学、结构和遗传学方法。 抗体将用于研究 血影蛋白在细胞培养和发育过程中， 鉴定表达果蝇血影蛋白DNA序列。 克隆 这些DNA序列将被用来绘制物理位置 果蝇基因图谱上血影蛋白的编码序列 经典的遗传分析和体外突变体生产将 然后执行。 变种人将被用来调查 血影蛋白在体内的功能，并确定 完全没有功能性血影蛋白。 克隆人也将 结合反义抑制技术， 表达以抑制培养细胞中血影蛋白的表达， 我们可以探测功能性血影蛋白缺乏的影响， 突变血影蛋白在培养细胞中表达的后果。 Spectrin存在于几乎所有的人体细胞中，但健康 血影蛋白表达缺陷的后果仅限于 在红细胞中探索。 拟议的实验应 提供了一个深入了解spectrin的作用，在非 红系细胞