Screening for bioactive small molecules by in vivo monitoring of luciferase-based reporter gene expression in Arabidopsis thaliana.
Screening for bioactive small molecules by in vivo monitoring of luciferase-based reporter gene expression in Arabidopsis thaliana.
复制标题
通过体内监测拟南芥中基于荧光素酶的报告基因表达来筛选生物活性小分子。
DOI：
10.1007/978-1-62703-592-7_3
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发表时间：
2014
影响因子：
--
通讯作者：
E. Kombrink
中科院分区：
文献类型：
--
作者：
C. Meesters;E. Kombrink
化学遗传学是一种利用生物活性小分子作为实验工具来剖析生物过程的科学策略。自然界中存在的生物活性化合物代表了结构的巨大多样性，这些结构潜在地可用作生化途径、运输过程、调控网络或发育程序的激活剂或抑制剂。鉴定生物活性小分子的筛选方法可以有很大的不同，从表型改变的视觉评估到定量生物特征如酶活性。在这里，我们描述了一个通用的方法，允许识别化合物调节拟南芥幼苗中的报告基因的表达。选择基于双磷酸酶的报告系统的优点在于，它允许以半定量的方式对报告基因活性进行体内成像，而不影响植物的生存力。我们选择了一个拟南芥线窝藏的茉莉酸诱导型LOX 2启动子的控制下的荧光素酶报告基因表达的激活剂或抑制剂进行筛选。概述的测定条件可以容易地应用于含有其他报告基因的拟南芥品系。因此，筛选影响不同信号传导途径和/或表型应答的小分子是可能的。
Chemical genetics is a scientific strategy that utilizes bioactive small molecules as experimental tools to dissect biological processes. Bioactive compounds occurring in nature represent an enormous diversity of structures that potentially can be used as activators or inhibitors of biochemical pathways, transport processes, regulatory networks, or developmental programs. Screening methods to identify bioactive small molecules can vary greatly, ranging from visual evaluation of phenotypic alterations to quantifying biometric traits such as enzyme activities. Here, we describe a general methodology that permits identification of compounds modulating the expression of reporter genes in Arabidopsis thaliana seedlings. The selection of luciferase-based reporter systems has the advantage that it allows in vivo imaging of reporter gene activity in a semiquantitative manner without affecting plant viability. We chose an Arabidopsis line harboring the luciferase reporter under the control of the jasmonate-inducible LOX2 promoter to screen for either activators or inhibitors of gene expression. The outlined assay conditions can readily be applied to Arabidopsis lines containing other reporter genes. Thereby screening for small molecules affecting different signaling pathways and/or phenotypic responses is possible.
DOI：
10.1073/pnas.0806324105
发表时间：
2008-09-30
影响因子：
11.1
作者：
Savaldi-Goldstein, Sigal;Baiga, Thomas J.;Chory, Joanne
通讯作者：
Chory, Joanne
影响因子：
27.5
作者：
Gendron, Joshua M.;Haque, Asif;Wang, Zhi-Yong
通讯作者：
Wang, Zhi-Yong