INTERSPECIFIC VARIATION OF ADH EXPESSION IN DROSOPHILA

果蝇 ADH 表达的种间变异

• 批准号: 3290265
• 负责人: CATHY C LAURIE
• 金额: $ 14.18万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-07-01 至 1991-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3290265
• 关键词: Drosophilidae; alcohol dehydrogenase; developmental genetics; drug tolerance; evolution; gene expression; genetic manipulation; genetic mapping; genetic recombination; genotype; larva; mature animal; molecular cloning; mutant; nucleic acid sequence; site directed mutagenesis; species difference

The goal of this project is to investigate the molecular basis of variation in the pattern and level of alcohol dehydrogenase (Adh) expression among the eight melanogaster subgroup species of Drosophila. The proposed work is both descriptive and experimental. In the descriptive phase, variation in the developmental profiles and tissue-distribution of ADH level and in the alcohol tolerance/utilization of larvae and adults will be analyzed for all eight species. A detailed analysis of the melanogaster/simulans species pair at the ADH-protein and ADH-RNA levels will be made in order to further investigate previous reports of a marked difference in the pattern of Adh expression. The experimental phase will utilize P element mediated transformation in order to determine the molecular basis of the melanogaster/simulans difference. First, reciprocal transfer of an Adh-containing DNA fragment between ADH-negative strains of each species will establish the extent to which the interspecific difference in Adh expression is due to nucleotide substitutions in the Adh region itself as opposed to background genotype differences. The precise molecular basis of any effect that maps to the Adh fragment will be determined by in vitro recombination and site-directed mutagenesis. Transformant stocks will also be analyzed to determine the contribution of variation in the Adh region to interspecific variation in alcohol tolerance/utilization. This strategy provides a direct approach to investigating the relative importance of structural versus regulatory variants in adaptive evolutionary change.

这个项目的目标是研究变异的分子基础 乙醇脱氢酶（Adh）表达的模式和水平， 果蝇的八个黑腹亚种。 拟议工作 既是描述性的也是实验性的。 在描述阶段，变异 ADH水平的发育概况和组织分布， 分析幼虫和成虫的酒精耐受性/利用率， 八个物种。 对黑腹/拟人的详细分析 将在ADH-蛋白和ADH-RNA水平上进行物种配对， 进一步调查以前关于模式明显不同的报告， Adh的表达。 实验阶段将利用P元件介导 转化，以确定的分子基础， melanogaster/simulans差异。 第一，相互转让 各菌种ADH阴性菌株间含ADH的DNA片段 将确定在何种程度上种间差异， 表达是由于Adh区域本身的核苷酸取代， 与背景基因型差异相反。 精确的分子基础 映射到Adh片段的任何效应将通过体外 重组和定点诱变。 转型股也将 分析以确定Adh区域中的变化对 酒精耐受/利用的种间差异。 这一战略 提供了一个直接的方法来调查的相对重要性， 适应性进化变化中的结构变异与调节变异。