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监测缺氧诱导因子(HIF)和核因子κB(NFκB)活性的转录动力学。

Monitoring of transcriptional dynamics of HIF and NFκB activities.

作者信息

Cavadas Miguel A S, Cheong Alex

机构信息

Systems Biology Ireland, University College Dublin, Dublin, Ireland.

出版信息

Methods Mol Biol. 2014;1098:97-105. doi: 10.1007/978-1-62703-718-1_8.

Abstract

Genetic experiments over the last few decades have identified many regulatory proteins critical for DNA transcription. The dynamics of their transcriptional activities shape the differential expression of the genes they control. Here we describe a simple method, based on the secreted luciferase, to measure the activities of two transcription factors NFκB and HIF. This technique can effectively monitor dynamics of transcriptional events in a population of cells and be up-scaled for high-throughput screening and promoter analysis, making it ideal for data-demanding applications such as mathematical modelling.

摘要

过去几十年的遗传学实验已经鉴定出许多对DNA转录至关重要的调控蛋白。它们转录活性的动态变化塑造了其所控制基因的差异表达。在此,我们描述一种基于分泌型荧光素酶的简单方法,用于测量两种转录因子NFκB和HIF的活性。该技术能够有效监测细胞群体中转录事件的动态变化,并且可以扩大规模用于高通量筛选和启动子分析,使其成为数学建模等需要大量数据的应用的理想选择。

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