Fu Yu, Xiao Wei
Department of Microbiology and Immunology, University of Saskatchewan, Saskatoon, Canada.
Methods Mol Biol. 2006;313:257-64. doi: 10.1385/1-59259-958-3:257.
Study of gene expression can be facilitated by using a reporter gene assay. Instead of directly measuring the level of target gene mRNA, one can clone the promoter region of the gene of interest in front of a reporter gene and measure the reporter gene expression as a reflection of the expression of the gene of interest. We describe a simple lacZ-fusion system to measure the activity of the reporter gene product beta-galactosidase. Different strategies of making the fusion construct and their applications are also discussed. This method is particularly useful to dissect the promoter region of the gene of interest and is also used in other experimental protocols such as the yeast two-hybrid analysis.
使用报告基因检测法有助于基因表达的研究。无需直接测量靶基因mRNA的水平,人们可以将感兴趣基因的启动子区域克隆到报告基因之前,并测量报告基因的表达,以此反映感兴趣基因的表达情况。我们描述了一种简单的lacZ融合系统,用于测量报告基因产物β-半乳糖苷酶的活性。还讨论了构建融合体的不同策略及其应用。该方法对于剖析感兴趣基因的启动子区域特别有用,也用于其他实验方案,如酵母双杂交分析。