Repressible Gal4-UAS expression systems for Drosphila

果蝇可抑制 Gal4-UAS 表达系统

• 批准号: 6353273
• 负责人: JAMES P MOHLER
• 金额: $ 14.32万
• 依托单位: BARNARD COLLEGE
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6353273
• 关键词: DNA binding protein; Drosophilidae; bacterial proteins; developmental genetics; fusion gene; gene expression; gene induction /repression; genetic models; genetic techniques; immunocytochemistry; in situ hybridization; method development; model design /development; polymerase chain reaction; transcription factor; transfection

DESCRIPTION (provided by applicant): One of the most useful tools for the study of developmental genetics in Drosophila is the Ga14-UAS modular expression system. This system allows the use of a wide variety of Ga14 expression lines to express transgenes in diverse tissues. The purpose of the research proposed here is to modify this system with a second level of negative control to allow the activation of a transgene by Ga14 in a set of Drosophila tissues to be tailored to a specific subset of tissues. Repressors based on the bacterial DNA-binding proteins, TetR and LexA, will be made by fusing their DNA-binding domains with repression domains from the Drosophila proteins Hairy and Groucho. The TetR-based repressor function should be regulated by administration of tetracycline, allowing the modulation of Ga14-mediated activation at different developmental stages. The LexA-based repressor will be regulated transcriptionally, such that only tissues with the Ga14-activator but not the LexA-repressor will be capable of expressing a transgene.

描述（由申请人提供）：研究中最有用的工具之一 果蝇发育遗传学的一个重要特征是Ga 14-UAS模块表达 系统该系统允许使用各种各样的Ga 14表达系 在不同的组织中表达转基因。提出的研究目的 这里是用第二水平的阴性对照来修改该系统， 在一组果蝇组织中通过Ga 14激活转基因， 针对特定的组织亚群定制。 基于细菌DNA结合蛋白TetR和莱克萨的抑制子将被 通过将它们的DNA结合结构域与抑制结构域融合， 果蝇蛋白质Hairy和Groucho。基于TetR的阻遏物功能应该 通过给予四环素进行调节， 在不同发育阶段的Ga 14介导的激活。基于LexA 阻遏物将在转录上受到调节，使得只有具有阻遏物的组织才能被激活。 Ga 14-激活物而不是LexA-阻遏物将能够表达 转基因。