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一个鉴定果蝇翅膀生长和形态形成所需基因的功能获得性筛选。

A gain-of-function screen identifying genes required for growth and pattern formation of the Drosophila melanogaster wing.

机构信息

Centro de Biología Molecular "Severo Ochoa" (CBMSO), Consejo Superior de Investigaciones Científicas and Universidad Autónoma de Madrid, Cantoblanco, Madrid 28049, Spain.

出版信息

Genetics. 2009 Nov;183(3):1005-26. doi: 10.1534/genetics.109.107748. Epub 2009 Sep 7.

Abstract

The Drosophila melanogaster wing is a model system for analyzing the genetic control of organ size, shape, and pattern formation. The formation of the wing involves a variety of processes, such as cell growth, proliferation, pattern formation, and differentiation. These developmental processes are under genetic control, and many genes participating in specific aspects of wing development have already being characterized. In this work, we aim to identify novel genes regulating wing growth and patterning. To this end, we have carried out a gain-of-function screen generating novel P-UAS (upstream activating sequences) insertions allowing forced gene expression. We produced 3,340 novel P-UAS insertions and isolated 300 that cause a variety of wing phenotypes in combination with a Gal4 driver expressed exclusively in the central domain of the presumptive wing blade. The mapping of these P-UAS insertion sites allowed us to identify the gene that causes the gain-of-function phenotypes. We show that a fraction of these phenotypes are related to the induction of cell death in the domain of ectopic gene expression. Finally, we present a preliminary characterization of a gene identified in the screen, the function of which is required for the development of the L5 longitudinal vein.

摘要

黑腹果蝇的翅膀是分析器官大小、形状和模式形成的遗传控制的模型系统。翅膀的形成涉及多种过程,如细胞生长、增殖、模式形成和分化。这些发育过程受遗传控制,许多参与翅膀发育特定方面的基因已经被描述。在这项工作中,我们旨在鉴定调节翅膀生长和模式形成的新基因。为此,我们进行了一个正向功能筛选,生成了新的 P-UAS(上游激活序列)插入,允许强制基因表达。我们产生了 3340 个新的 P-UAS 插入,并分离了 300 个与仅在翼片前体中央域表达的 Gal4 驱动器结合导致各种翅膀表型的插入。这些 P-UAS 插入位点的映射使我们能够鉴定导致正向功能表型的基因。我们表明,这些表型的一部分与异位基因表达域中的细胞死亡诱导有关。最后,我们介绍了在筛选中鉴定的一个基因的初步特征,该基因的功能对于 L5 纵向静脉的发育是必需的。

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