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Genetic and molecular analysis of mutations in Drosophila melanogaster that lead to differences in expression of microRNA mir-92a and underlie phenotypic evolution

对果蝇中导致 microRNA mir-92a 表达差异和表型进化基础的突变进行遗传和分子分析

基本信息

批准号：
241455725
负责人：
Dr. Sebastian Kittelmann
金额：
--
依托单位：
Evolution of Animal Development and Morphology Group
依托单位国家：
德国
项目类别：
Research Fellowships
财政年份：
2013
资助国家：
德国
起止时间：
2012-12-31 至 2016-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/241455725?language=en
关键词：
Genetic molecular analysis mutations Drosophila

项目摘要

It is a goal of evolutionary biology to link the variation of morphological traits to the underlying genetic variation. A trait that differs between different Drosophila species but is usually constant within each species is the size of the naked valley, a naked cuticle patch on the second leg. However, several populations of Drosophila melanogaster show size differences of the naked valley. The underlying genetic variation has been mapped to a locus containing a microRNA, mir-92a, which is located in an intron of the protein-coding gene jigr1. mir-92a post-transcriptionally regulates shavenoid (sha), a gene involved in naked valley development. I will first examine the possibility of jigr1 or a second protein-coding gene, Npl4, being involved in the development of the naked valley, as well. Differences in expression of mir-92a in the different Drosophila strains are thought to underlie the naked valley size variation by subsequent differential regulation of sha. Differential expression of mir-92a is likely to be facilitated by mutations in an enhancer. I will analyse whether expression of mir-92a and jigr1 is correlated and thus controlled by the same cis-regulatory elements. In order to identify the size-determining element, I will use the GAL4-UAS system to express GFP as well as mir-92a under control of candidate enhancers from strains with large and small naked valleys, respectively, in the genetic background of a small naked valley. I will identify the specific mutations responsible for the size variation by stepwise altering the enhancer for development of a large naked valley towards the identity of a strain with a small naked valley by PCR-based mutagenesis. The results of these experiments will facilitate the identification of the mutations underlying phenotypic variation of the naked valley within Drosophila melanogaster.

将形态性状的变异与潜在的遗传变异联系起来是进化生物学的目标之一。裸谷的大小在不同的果蝇物种之间有所不同，但在每个物种中通常是恒定的，裸谷是第二条腿上的一块裸露的角质层。然而，黑腹果蝇的几个种群显示出裸谷的大小差异。潜在的遗传变异已被定位到一个包含microRNA mir-92a的基因座，该基因位于蛋白质编码基因jgr1的一个内含子中。Mir-92a转录后调控与裸谷发育有关的shavenid基因。我将首先研究jgr1或第二个编码蛋白质的基因npl4是否也参与裸谷的发展。不同果蝇品系mir-92a表达的差异被认为是随后对SHA的差异调控导致裸谷大小差异的原因。增强子的突变可能促进mir-92a的差异表达。我将分析mir-92a和jgr1的表达是否相关，从而受到相同的顺式调控元件的控制。为了鉴定大小决定元件，我将使用GAL4-UAS系统在一个小裸谷的遗传背景下，分别从大裸谷和小裸谷菌株的候选增强子控制下表达GFP和mir-92a。我将通过基于聚合酶链式反应的突变，逐步改变大裸谷发育的增强子，使其与小裸谷的菌株相同，从而鉴定导致大小变化的特定突变。这些实验的结果将有助于鉴定果蝇裸谷表型变异的潜在突变。