Establishment of a newly improved detection system for NF-κB activity
Establishment of a newly improved detection system for NF-κB activity
复制标题
DOI：
10.1016/j.imlet.2007.02.007
复制
发表时间：
2007-04-15
影响因子：
4.4
通讯作者：
Hatakeyama, Shigetsugu
中科院分区：
文献类型：
--
作者：
Matsuda, Mayuko;Tsukiyama, Tadasuke;Hatakeyama, Shigetsugu
转录因子核因子 kappa B (NF-kappa B) 在细胞凋亡、炎症和肿瘤发生中发挥作用。对于生物学和医学研究来说，了解感兴趣的蛋白质何时在细胞中被激活非常重要，从而建立荧光素酶/EGFP 检测来监测转录因子的激活。在这里，我们描述了一种改进的 NF-kappa B 报告系统，即 NF-kappa B 激活的转基因 (NAT) 系统，可以高灵敏度和特异性地检测 NF-kappa B 信号传导。 NAT 系统由大量拷贝的 NF-kappa B 共有序列和源自小鼠白细胞介素 2 (IL-2) 基因的最小启动子组成。此外，我们还生成了具有稳定或不稳定荧光素酶/EGFP 蛋白的 NAT 系统。稳定和不稳定类型的荧光素酶/EGFP分别适合分析NF-kappa B信号的积累和实时活性。我们的研究结果表明，NAT 系统对于使用细胞或动物进行 NF-kappa B 信号传导的体内成像是有效的。 (c) 2007 Elsevier B.V. 保留所有权利。
The transcription factor nuclear factor-kappa B (NF-kappa B) plays roles in apoptosis, inflammation and oncogenesis. It is important for biological and medical research to understand when proteins of interest are activated in cells, leading to the establishment of a luciferase/EGFP assay to monitor the activation of transcription factors. Here, we describe an improved reporter system for NF-kappa B, the NF-kappa B-activated transgene (NAT) system that can detect NF-kappa B signalling with high sensitivity and specificity. The NAT system consists of large copy numbers of NF-kappa B consensus sequence and a minimal promoter derived from the mouse interleukin-2 (IL-2) gene. Furthermore, we generated NAT systems with stable or unstable luciferase/EGFP proteins. Stable and unstable types of luciferase/EGFP are suitable for analyzing the accumulation of and the real-time activity of NF-kappa B signal, respectively. Our findings suggest that the NAT system is effective for in vivo imaging of NF-kappa B signalling using cells or animals. (c) 2007 Elsevier B.V. All rights reserved.