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Functional Analysis of Genes Differentially Expressed in the Drosophila Wing Disc: Role of Transcripts Enriched in the Wing Region

机译:果蝇翼盘中差异表达基因的功能分析:翼区中丰富的转录本的作用

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摘要

Differential gene expression is the major mechanism underlying the development of specific body regions. Here we assessed the role of genes differentially expressed in the Drosophila wing imaginal disc, which gives rise to two distinct adult structures: the body wall and the wing. Reverse genetics was used to test the function of uncharacterized genes first identified in a microarray screen as having high levels of expression in the presumptive wing. Such genes could participate in elaborating the specific morphological characteristics of the wing. The activity of the genes was modulated using misexpression and RNAi-mediated silencing. Misexpression of eight of nine genes tested caused phenotypes. Of 12 genes tested, 10 showed effective silencing with RNAi transgenes, but only 3 of these had resulting phenotypes. The wing phenotypes resulting from RNAi suggest that CG8780 is involved in patterning the veins in the proximal region of the wing blade and that CG17278 and CG30069 are required for adhesion of wing surfaces. Venation and apposition of the wing surfaces are processes specific to wing development providing a correlation between the expression and function of these genes. The results show that a combination of expression profiling and tissue-specific gene silencing has the potential to identify new genes involved in wing development and hence to contribute to our understanding of this process. However, there are both technical and biological limitations to this approach, including the efficacy of RNAi and the role that gene redundancy may play in masking phenotypes.
机译:差异基因表达是特定身体部位发育的主要机制。在这里,我们评估了在果蝇翅假想盘中差异表达的基因的作用,这导致了两个不同的成年结构:体壁和翅。反向遗传学用于测试未鉴定基因的功能,该基因最初在微阵列筛选中鉴定为在推测翼中具有高表达水平。这样的基因可以参与阐述机翼的特定形态特征。基因的活性是通过错误表达和RNAi介导的沉默来调节的。测试的9个基因中有8个的错误表达导致了表型。在测试的12个基因中,有10个显示出对RNAi转基因的有效沉默,但其中只有3个具有最终的表型。 RNAi引起的机翼表型表明,CG8780参与了机翼叶片近端区域的静脉构图,而CG17278和CG30069是机翼表面粘附所必需的。机翼表面的通风和附着是机翼发育所特有的过程,可在这些基因的表达和功能之间提供相关性。结果表明,表达谱分析和组织特异性基因沉默相结合,有可能识别参与机翼发育的新基因,从而有助于我们对这一过程的理解。但是,这种方法在技术和生物学上都存在局限性,包括RNAi的功效以及基因冗余可能在掩盖表型中发挥的作用。

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