INTERSPECIFIC VARIATION OF ADH EXPESSION IN DROSOPHILA

这个项目的目标是研究变异的分子基础。 乙醇脱氢酶(ADH)的表达模式和水平 果蝇的八种黑腹果蝇亚群。拟议中的工作 既是描述性的，也是实验性的。在描述阶段，变体 在ADH水平和IN的发育概况和组织分布中 将分析幼虫和成虫的酒精耐受性/利用 全部八个物种。黑猩猩/模拟者的详细分析 将在ADH-蛋白质和ADH-RNA水平上进行物种配对，以便 进一步调查之前关于模式明显不同的报道 ADH的表达。实验阶段将利用P元素介导 转化，以确定分子基础的 黑猩猩/拟人猿的区别。第一，互惠转让 各种ADH阴性菌株间含ADH的DNA片段 将确定ADH种间差异的程度 表达是由于adh区本身的核苷酸替换所致 而不是背景基因差异。分子基础的精确度 任何映射到adh片段的作用都将通过体外实验来确定。 重组和定点突变。变革股也将 被分析以确定ADH区域的变异对 酒精耐受性/利用的种间差异。这一战略 提供了一种直接方法来调查 适应性进化变化中的结构变量与调控变量。

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.