HOMOLOG OF DROSPHILA RDGB

果蝇 RDGB 的同源物

• 批准号: 2882938
• 负责人: JINGHUA T CHANG
• 金额: $ 11.78万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-03-01 至 2000-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2882938
• 关键词: Drosophilidae; SDS polyacrylamide gel electrophoresis; apoptosis; calcium binding protein; electron microscopy; electroretinography; enzyme activity; gene expression; gene mutation; genetically modified animals; immunocytochemistry; in situ hybridization; laboratory mouse; light microscopy; northern blottings; phosphatidylinositols; polymerase chain reaction; protein structure function; retina degeneration; single cell analysis; vision; visual photoreceptor; visual phototransduction; western blottings; yeast two hybrid system

DESCRIPTION (Adapted from applicant's abstract): The long-term objective of this project is to study the biological function of the mammalian ortholog of Drosophila rdgB (mrdgB) in the visual process. Mutations of rdgB cause light-enhanced retinal degeneration in flies. The MrdgB and RdgB proteins share high degrees of structural and functional conservation. When mrdgB is expressed in rdgB mutant flies, it suppresses the light-enhanced retinal degeneration and restores normal ERGs. This finding is surprising given the significant morphological and biochemical differences between fly and vertebrate eyes, and suggests the possibility of functional similarities between the species that were not previously appreciated. The biological functions of either rdgB or mrdgB are not yet known. In order to begin to understand the role of mrdgB in the mammalian visual system, several experiments are proposed. First, the developmental expression pattern of mrdgB in the mouse eye will be characterized. Also, because rdgB exhibits phosphatidylinositol transferase (PIT) and calcium-binding activities, the MrdgB protein will be assayed to determine whether it also possesses these biochemical activities in vitro. The most direct method to assess mrdgB function in the mammalian retina is to disrupt its function. Therefore, the main focus of this project will be to generate mrdgB knock-out mice. This will be achieved by targeted gene disruption in murine embryonic stem cells. Another effective way to study the function of a protein is to isolate and characterize proteins with which it interacts. To achieve this, the yeast two-hybrid system will be used to characterize proteins that interact with MrdgB. Hopefully, these studies will provide new insights into the biology of vision and increase our understanding of human retinal degeneration.

描述（根据申请人的摘要改编）： 该项目是研究哺乳动物直系同源物的生物学功能 在视觉过程中果蝇RDGB（MRDGB）的摄入。 RDGB原因的突变 苍蝇的轻度视网膜变性。 MRDGB和RDGB蛋白 共享高度的结构和功能保护。 当mrdgb是时 在RDGB突变蝇中表达，它抑制了视网膜的光。 变性并恢复正常的ERG。 考虑到这一发现令人惊讶 苍蝇与 脊椎动物的眼睛，并提出功能相似性的可能性 在以前未被理解的物种之间。 RDGB或MRDGB的生物学功能尚不清楚。 在 为了开始了解MRDGB在哺乳动物视觉中的作用 系统，提出了一些实验。 首先，发展性 将表征MRDGB在小鼠眼中的表达模式。 还， 因为RDGB表现出磷脂酰肌醇转移酶（PIT）和 钙结合活性，将分析MRDGB蛋白以确定 它是否还在体外具有这些生化活性。 最多 评估哺乳动物视网膜中MRDGB功能的直接方法是破坏 它的功能。 因此，该项目的主要重点是生成 MRDGB淘汰小鼠。 这将通过靶向基因破坏来实现 鼠类胚胎干细胞。 研究功能的另一种有效方法 蛋白质是孤立和表征与之相互作用的蛋白质的。 为此，将使用酵母双杂交系统来表征 与MRDGB相互作用的蛋白质。 希望这些研究能够提供 对视觉生物学的新见解，并提高我们对 人类视网膜变性。