Southall Tony D, Elliott David A, Brand Andrea H
The Gurdon Institute and Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, United Kingdom.
CSH Protoc. 2008 Jul 1;2008:pdb.top49. doi: 10.1101/pdb.top49.
INTRODUCTIONThe generation of gain-of-function phenotypes by ectopic expression of known genes provides a powerful complement to the genetic approach, in which genes are studied or identified through mutations that generally reduce or eliminate gene function. The GAL4 system is a method for ectopic gene expression that allows the selective activation of any cloned gene in a wide variety of tissue- and cell-specific patterns. A key advantage of the system is the separation of the GAL4 protein from its target gene in distinct transgenic lines, which ensures that the target gene is silent until the introduction of GAL4. Recent modifications and adaptations of the GAL4 system to make the system inducible have further expanded its scope, enabling greater temporal control over the activity of GAL4. There are now large resources for the community, including thousands of GAL4 lines and a wide selection of reporter lines. Here we present an overview of the GAL4 system, highlighting recent developments and discussing methods for generating and analyzing transgenic flies for GAL4-mediated ectopic expression.
引言
通过已知基因的异位表达产生功能获得性表型,为遗传学方法提供了有力补充。在遗传学方法中,基因是通过通常会降低或消除基因功能的突变来进行研究或鉴定的。GAL4系统是一种用于异位基因表达的方法,它允许以多种组织和细胞特异性模式选择性激活任何克隆基因。该系统的一个关键优势是在不同的转基因品系中,GAL4蛋白与其靶基因分离,这确保了靶基因在引入GAL4之前保持沉默。GAL4系统最近的改进和调整使其具有诱导性,进一步扩大了其应用范围,能够对GAL4的活性进行更好的时间控制。目前有大量资源可供研究群体使用,包括数千个GAL4品系和多种报告基因品系。在这里,我们概述了GAL4系统,重点介绍了近期的进展,并讨论了用于生成和分析GAL4介导的异位表达转基因果蝇的方法。