Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge CB2 3DY, United Kingdom.
G3 (Bethesda). 2012 Jan;2(1):79-82. doi: 10.1534/g3.111.001321. Epub 2012 Jan 1.
Complex spatial and temporal regulation of gene activity is fundamental to development and homeostasis. The ability to decipher the DNA sequences that accurately coordinate gene expression is, therefore, of primary importance. One way to assess the functions of DNA elements entails their fusion to fluorescent reporter genes. This powerful approach makes it possible to visualize their regulatory capabilities when reintroduced into the developing animal. Transgenic studies in Drosophila have recently advanced with the introduction of site-specific, ΦC31 integrase-mediated approaches. However, most existing Drosophila reporter vectors are not compatible with this new approach and have become obsolete. Here we describe a new series of fluorescent reporter vectors optimized for use with ΦC31 transgenesis. By using these vectors to generate a set of Notch reporter fly lines, we demonstrate their efficacy in reporting the function of gene regulatory elements.
基因活性的复杂时空调控对发育和内稳态至关重要。因此,破译能够准确协调基因表达的 DNA 序列至关重要。评估 DNA 元件功能的一种方法是将其与荧光报告基因融合。这种强大的方法使得在将其重新引入发育中的动物时可以可视化其调节能力。最近,随着定点、ΦC31 整合酶介导方法的引入,果蝇的转基因研究取得了进展。然而,大多数现有的果蝇报告载体与这种新方法不兼容,已经过时。在这里,我们描述了一系列新的荧光报告载体,这些载体经过优化可与 ΦC31 转基因技术一起使用。通过使用这些载体生成一组 Notch 报告蝇系,我们证明了它们在报告基因调控元件功能方面的功效。