A versatile ΦC31 based reporter system for measuring AP-1 and Nrf2 signaling in Drosophila and in tissue culture.
A versatile ΦC31 based reporter system for measuring AP-1 and Nrf2 signaling in Drosophila and in tissue culture.
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一种基于多功能φC31的记者系统，用于测量果蝇和组织培养中的AP-1和NRF2信号传导。
DOI：
10.1371/journal.pone.0034063
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Bohmann D
中科院分区：
文献类型：
--
作者：
Chatterjee N;Bohmann D
本文描述了一个转录报告基因系统的构建和特性，该系统用于监测完整的果蝇、解剖组织或培养细胞中信号通路的活性和基因调控机制。利用ΦC31整合酶以定点的方式将这些报告基因整合到果蝇胚系中。该策略避免了可变位置效应，并确保了低基准值活动和高信号响应性。此外，确定的整合位点还使实验者能够比较同一生物体中不同报告基因的活性。报告构建体具有模块化设计，以促进启动子元件(合成转录因子结合位点或自然调节序列)、报告基因(EGFP或DsRed.T4)和基因组整合位点的结合。利用该系统分析和比较了AP-1和Nrf2两种逆境诱导转录因子的活性和信号响应谱。为了补充转基因报告蝇系，开发了组织培养实验，其中相同的合成ARE和TrE元件控制萤火虫荧光素酶的表达。
This paper describes the construction and characterization of a system of transcriptional reporter genes for monitoring the activity of signaling pathways and gene regulation mechanisms in intact Drosophila, dissected tissues or cultured cells. Transgenic integration of the reporters into the Drosophila germline was performed in a site-directed manner, using ΦC31 integrase. This strategy avoids variable position effects and assures low base level activity and high signal responsiveness. Defined integration sites furthermore enable the experimenter to compare the activity of different reporters in one organism. The reporter constructs have a modular design to facilitate the combination of promoter elements (synthetic transcription factor binding sites or natural regulatory sequences), reporter genes (eGFP, or DsRed.T4), and genomic integration sites. The system was used to analyze and compare the activity and signal response profiles of two stress inducible transcription factors, AP-1 and Nrf2. To complement the transgenic reporter fly lines, tissue culture assays were developed in which the same synthetic ARE and TRE elements control the expression of firefly luciferase.
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影响因子：
4
作者：
Hess, J;Angel, P;Schorpp-Kistner, M
通讯作者：
Schorpp-Kistner, M
影响因子：
3.3
作者：
Bateman, Jack R.;Lee, Anne M.;Wu, C. -ting
通讯作者：
Wu, C. -ting
影响因子：
1.5
作者：
Boy, Aurelia L.;Zhai, Zongzhao;Lohmann, Ingrid
通讯作者：
Lohmann, Ingrid
影响因子：
14.9
作者：
Arziman Z;Horn T;Boutros M
通讯作者：
Boutros M
DOI：
10.1073/pnas.94.11.5826
发表时间：
1997-05-27
影响因子：
11.1
作者：
Huang, CS;Ma, WY;Dong, ZG
通讯作者：
Dong, ZG