Scheer N, Campos-Ortega J A
Institut für Entwicklungsbiologie, Universität zu Köln, 50923, Cologne, Germany.
Mech Dev. 1999 Feb;80(2):153-8. doi: 10.1016/s0925-4773(98)00209-3.
The most common way to analyze the function of cloned genes in zebrafish is to misexpress the gene product or an altered variant of it by mRNA injection. However, mRNA injection has several disadvantages. The GAL4-UAS system for targeted gene expression allows one to overcome some of these disadvantages. To test the GAL4-UAS system in zebrafish, we generated two different kinds of stable transgenic lines, carrying activator and effector constructs, respectively. In the activator lines the gene for the yeast transcriptional activator GAL4 is under the control of a given promoter, while in the effectors the gene of interest is fused to the sequence of the DNA-binding motif of GAL4 (UAS). Crosses of animals from the activator and effector lines show that effector genes are transcribed with the spatial pattern of the activators. This work smoothes the way for a novel method of misexpression of gene products in zebrafish in order to analyze the function of genes in developmental processes.
在斑马鱼中分析克隆基因功能最常见的方法是通过注射mRNA使基因产物或其变异体错误表达。然而，mRNA注射存在几个缺点。用于靶向基因表达的GAL4-UAS系统能够克服其中一些缺点。为了在斑马鱼中测试GAL4-UAS系统，我们分别构建了携带激活子和效应子构建体的两种不同类型的稳定转基因品系。在激活子品系中，酵母转录激活因子GAL4的基因受特定启动子的控制，而在效应子品系中，目的基因与GAL4的DNA结合基序序列（UAS）融合。激活子品系和效应子品系的动物杂交表明，效应基因按照激活子的空间模式进行转录。这项工作为一种在斑马鱼中错误表达基因产物以分析基因在发育过程中功能的新方法铺平了道路。
