ROLE OF THE SWALLOW GENE IN DROSOPHILA DEVELOPMENT

燕子基因在果蝇发育中的作用

• 批准号: 3299758
• 负责人: EDWIN C STEPHENSON
• 金额: $ 4.62万
• 依托单位: UNIVERSITY OF ALABAMA IN TUSCALOOSA
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-12-01 至 1992-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3299758
• 关键词: Drosophilidae; RNA biosynthesis; binding proteins; complementary DNA; cytoskeletal proteins; developmental genetics; early embryonic stage; egg /ovum; endonuclease; fluorescence microscopy; gene expression; gene mutation; gene redundancy; genes; genetic manipulation; genetic transcription; immunoprecipitation; messenger RNA; nonmammalian vertebrate embryology; nucleic acid sequence; oogenesis; physical separation; protein biosynthesis; pseudogenes

The rationale for experiments proposed in this application begins with the observation that the mRNA of a Drosophila material effect gene, bicoid, is localized at the anterior end of oocytes and early embryos. Further, two other maternal effect genes, swallow and exuperantia, are necessary for this localization. I propose to determine the function of the swallow gene in this process, and parameters of the bicoid message localization process itself. The localization of determinants in animal eggs is of widespread occurrence, and information gained with respect to the function of the swallow gene in Drosophila should be of fundamental importance in understanding developmental processes. This application includes the following experiments: (1) We will determine the significance of an unusual feature of the swallow gene region, the presence of two long direct repeats. We would like to know whether one or both repeats encode swallow function. (2) We will determine the time of synthesis and persistence of the swallow protein, and its location in the egg chamber and embryo at gross and subcellular levels. We will look at swallow protein distribution in oocytes and embryos from wildtype, swallow- and exuperantia- mothers. (3) We will examine the organization of the cytoskeleton in swallow- egg chambers and embryos, testing aspects of the idea that the swallow+ gene product has a role in cytoskeletal organization. (4) We will determine the effects of inhibitors of cytoskeletal organization on the localization of bicoid mRNA and swallow protein. (5) We will characterize the molecular associations of swallow protein and bicoid message in Drosophila oocytes and embryos by fractionation, immunoprecipitation and in vitro binding experiments.

本申请中提出的实验的基本原理始于 观察到果蝇材料效应基因Bicoid的mRNA是 位于卵母细胞和早期胚胎的前端。 此外，两个 其他母体效应基因，吞咽和陶器是必需的 这个本地化。 我建议确定燕子基因的功能 在此过程中，Bicoid消息定位过程的参数 本身。 动物卵中的决定因素的定位是广泛的 发生，并获得有关该功能的信息 果蝇中的吞咽基因在 了解发展过程。 该应用程序包括以下实验： （1）我们将确定燕子不寻常特征的重要性 基因区域，存在两个长直接重复。 我们想 知道一个或两个重复编码吞咽函数。 （2）我们将确定燕子的合成时间和持久性 蛋白质及其位于鸡蛋室和胚胎的位置 亚细胞水平。 我们将查看吞咽蛋白分布 来自野生型，燕子和散型母亲的卵母细胞和胚胎。 （3）我们将检查燕子中的细胞骨架的组织 腔室和胚胎，测试吞咽+基因的想法的方面 产品在细胞骨架组织中起作用。 （4）我们将确定细胞骨架抑制剂的影响 关于双子mRNA和吞咽蛋白的定位组织。 （5）我们将表征燕子蛋白和 果蝇卵母细胞和胚胎中的双聚体信息，通过分馏， 免疫沉淀和体外结合实验。