REGULATION OF CUTICLE GENE EXPRESSION IN DROSOPHILA

果蝇角质层基因表达的调控

基本信息

批准号：
3279907
负责人：
JAMES W FRISTROM
金额：
$ 8.13万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
1983
资助国家：
美国
起止时间：
1983-04-01 至 1988-03-31
项目状态：
已结题

项目摘要

Cuticle protein genes are comprised of 2 or 3 multigene families that are differentially expressed in response to the steroid hormone, 20-hydroxyecdysone (20HOE) during Drosophila development. Genes for larval cuticle proteins are expressed in the larval epidermis. Genes for pupal cuticle proteins, related to but distinct from the larval ones, are expressed in imaginal discs in prepupae. Expression of the pupas genes is induced in discs by 20HOE in vitro. Genomic clones containing a cluster of four 3rd instar cuticle protein genes have been recovered and characterized (Snyder et al., 1981) (35). Variants affecting the expression of these genes have been recovered. We propose to clone the genes for the pupal cuticle proteins and produce polyclonal and monoclonal antibody against these proteins. These reagents will be used to describe the molecular details of the onset of synthesis, in response to 20HOE, of cuticle proteins and their mRNAs in imaginal discs in vitro. We will systematically recover variants affecting the expression of larval and pupal cuticle genes. Both "cis"-acting and "distant regulators" of gene expression will be recovered and characterized genetically and at the DNA level. Pupal cuticle gene variants will be analyzed in discs cultured in vitro to determine the molecular nature of the alteration in expression using the cloned genes and the antibody to monitor synthesis. New information about the regulation of gene expression during development should result from these studies.

角质层蛋白基因由 2 或 3 个多基因家族组成，对类固醇激素的反应有差异表达，果蝇发育过程中的 20-羟基蜕皮酮 (20HOE)。幼虫基因角质层蛋白在幼虫表皮中表达。蛹的基因角质层蛋白与幼虫蛋白相关但又不同，在预蛹的成虫盘中表达。蛹基因的表达是 20HOE 体外在椎间盘中诱导。基因组克隆包含一簇四个三龄角质层蛋白基因已被恢复和表征（斯奈德等人，1981）（35）。影响这些表达的变体基因已被恢复。我们建议克隆蛹的基因角质层蛋白并产生多克隆和单克隆抗体这些蛋白质。这些试剂将用于描述分子角质层响应 20HOE 的合成开始细节体外成虫盘中的蛋白质及其 mRNA。我们将系统地恢复影响幼虫表达的变异体蛹角质层基因。基因的“顺式”作用和“远程调节器” 表达将被恢复并在遗传和 DNA 上进行表征等级。将在培养的圆盘中分析蛹角质层基因变异体外确定表达改变的分子性质使用克隆的基因和抗体来监测合成。新的有关发育过程中基因表达调控的信息应该是这些研究的结果。