MOLECULAR BIOLOGY OF VISION

这项研究的目的是为了深入了解 用于视觉系统中信号转导的机制， 结合分子遗传和生理学方法。 预计 从这些研究中获得的结果将有助于我们了解 生物学中感觉接受和信息处理的基础 系统. 在过去的两年半里，我们集中精力进行研究， 表征编码参与光转导的蛋白质的基因 级联。 我们分离了ninA基因（亲环素同源物）， 光感受器细胞特异性蛋白激酶C，一种果蝇同源物， 脊椎动物抑制蛋白和光感受器细胞特异性脱氢酶。 我们 我也继续我们的果蝇视蛋白和遗传分析， 在R1-R6中表达不同视蛋白的工程果蝇品系 感光细胞 这些实验使我们能够检查 各种视蛋白对特定功能特性的贡献 不同的感光细胞类型。 这项建议现在集中在我们的 这些基因家族中的两个：视蛋白和ninaA。 我们将：（1，2）确定光谱特异性的分子基础， 视蛋白分子(3)分离编码在大肠杆菌中表达的视蛋白的基因。 果蝇复眼的中央R8感光细胞类，和 (4)产生缺乏R7和R8视色素的突变体，和 从分子和生理学上描述它们。 我们还将：（5） 确定ninaA基因的表达位点，并产生抗体 对抗它的基因产物(6)产生过表达的转基因株系 ninaA，并在细菌（T7）中产生和表征ninaA蛋白 表达系统）和果蝇培养细胞（Schneider细胞）。 （七） 详细描述尼娜A的视觉生理和生物化学特征 苍蝇和确定野生型苍蝇的心理表型急性 并长期接受CsA治疗 (8)构造并表征热- 休克诱导型ninaA基因分离温度敏感型 ninaA. (9)分离ninaA基因家族的其他成员， 搜索推定的R7细胞特异性同源物。 最后，我们将在 详细描述了那些基于其表达模式的序列， 遗传和生化标准最值得进一步研究。

The aim of the research proposed in this grant is to gain insight into the mechanisms used for signal transduction in the visual system using a combined molecular, genetic and physiological approach. It is expected that the results obtained from these studies will help our understanding of the basis of sensory reception and information processing in biological systems. Over the past 2-1/2 years we have concentrated our research efforts characterizing genes encoding proteins involved in the phototransduction cascade. We isolated the ninA gene (cyclophilin-homologue), a photoreceptor-cell specific protein kinase C, a Drosophila homologue of vertebrate arrestin, and a photoreceptor-cell specific dehydrogenase. We have also continued our analysis of Drosophila opsins and genetically engineered Drosophila lines that express different opsins in the R1-R6 photoreceptor cells. These experiments allowed us to examine the contribution of the various opsins to the specific functional properties of the different photoreceptor cell types. This proposal now focus our research efforts on two of these gene families: opsins and ninaA. We will: (1,2) Determine the molecular basis of spectral specificities in the opsin molecule. (3) Isolate the gene(s) encoding the opsin expressed in the central R8 photoreceptor cell class of the Drosophila compound eye, and (4) generate mutants lacking the R7 and R8 visual pigments, and characterize them molecularly and physiologically. We will also: (5) Identify the sites of expression of the ninaA gene, and generate antibodies against its gene product. (6) Generate transgenic lines overexpressing ninaA, and produce and characterize the ninaA protein in bacteria (T7 expression system) and Drosophila culture cells (Schneider cells). (7) Characterize in detail the visual physiology and biochemistry of ninaA flies and determine the psychological phenotype of wild-type flies acutely and chronically treated with CsA. (8) Construct and characterize a heat- shock inducible ninaA gene an attempt to isolate temperature sensitive alleles of ninaA. (9) Isolate other members of the ninaA gene family, and search for a putative R7 cell-specific homolog. Finally, we will study in detail those sequences which based on their patterns of expression and genetic and biochemical criteria most warrant further investigation.