Analysis of the Inverse Effect in Drosophila

拟议工作的长期目标是了解 调节基因的性质和机制， 白色眼睛颜色基因的表达。 据信这 一种调节效应参与了大多数 结构基因的类型。 目前已知的四个逆 白色的调节器将被精确地定位，在遗传上， 细胞学上，通过X射线和P元素诱变作为前奏， 分子克隆 克隆其中两个基因将揭示它们的 性质，并提供手段，以解决分子机制， 通过序列分析、体外结合试验和 遗传剂量研究与转化副本。 四个 单独以及组合的反向调节器将 测试对发育中白色mRNA水平的影响 配置文件，以及从猩红色和 棕色，其在色素沉积中与白色配合。 双方都将 测试对白色启动子-报道基因构建体的影响， 检查W启动子的要求。 一个精致的功能 将进行白色启动子的分析以更好地定位 与四个监管机构互动所需的网站。 这 将通过删除和研究中心指导进行分析 启动子的诱变，然后将其置于驱动 乙醇脱氢酶结构基因 这些结构将是 测试，以确定四个逆调节器，并检查 这些地区的工作环境。 拟议的实验将有助于理解 逆调节现象及解决方法 这些调控基因与生物学特性之间的联系 与染色体剂量相关的现象如补偿， 逆效应、非整倍体综合征和位置效应 杂色。 //

The long range goal of the proposed work is to understand the nature and mechanism of regulatory genes that inversely affect the expression of the white eye color gene. It is believed that this type of regulatory effect is involved in the expression of most types of structural genes. The four currently known inverse regulators of white will be precisely localized, genetically and cytologically, via x-ray and P element mutagenesis as a prelude to molecular cloning. Cloning of two of these genes will reveal their nature and provide the means to address the molecular mechanism of action on white by sequence analysis, in vitro binding tests and genetic dosage studies with transformed copies. Each of the four inverse regulators, individually as well as in combinations, will be tested for an effect upon white mRNA levels in developmental profiles, as well as for a coordinate response from scarlet and brown, which cooperate with white in pigment deposition. Each will be tested for an effect on a white promoter-reporter construct to examine the requirement of the w promoter. A refined functional analysis of the white promoter will be conducted to better localize the sites required for interaction with the four regulators. This analysis will be conducted by deletion and site directed mutagenesis of the promoter, which will then be placed driving the alcohol dehydrogenase structural gene. These constructs will be tested to determine the four inverse regulators and examine the context within which these regions work. The proposed experiments will contribute to an understanding of the inverse regulation phenomenon and address whether there is a connection between such regulatory genes and the biological phenomena associated with chromosomal dosage such as compensation, the inverse effect, aneuploid syndromes, and position effect variegation.*** //