Single plasmids expressing human steroid hormone receptors and a reporter gene for use in yeast signaling assays.
Single plasmids expressing human steroid hormone receptors and a reporter gene for use in yeast signaling assays.
复制标题
DOI：
10.1016/j.plasmid.2009.11.003
复制
发表时间：
2010-03
期刊：
影响因子：
2.6
通讯作者：
Ludwig S
中科院分区：
文献类型：
--
作者：
Miller CA 3rd;Tan X;Wilson M;Bhattacharyya S;Ludwig S
本报告描述了设计用于表达六种人I型类固醇激素受体并检测信号传导活性的单质粒。这些稳定复制的质粒报告通过lacZ测定在面包酵母（酿酒酵母）配体诱导的转录激活。基于EC50值的比较，在表达这些质粒的酵母中激活信号传导所需的配体浓度跨越五个数量级。Radicicol，热休克蛋白90（HSP90）的直接抑制剂和类固醇激素受体信号的间接抑制剂，被用来确定该酵母报告系统的功能效用。根赤霉素对所有六种类固醇激素受体的信号传导的抑制作用相似，并且与较高浓度下发生的细胞毒性作用不同。这些酵母质粒提供了一个高通量的系统，用于比较类固醇激素受体信号转导的评估，并可能是有用的药理学或外源性活性的筛选。
Single plasmids designed to express the six human type I steroid hormone receptors and detect signaling activity are described in this report. These stably replicating plasmids reported ligand-induced transcriptional activation via lacZ assays in Baker's yeast (Saccharomyces cerevisiae). The ligand concentrations needed to activate signaling in yeast expressing these plasmids spanned five orders of magnitude as based on comparisons of EC50 values. Radicicol, a direct inhibitor of heat shock protein 90 (Hsp90) and an indirect inhibitor of steroid hormone receptor signaling, was used to determine the functional utility of this yeast reporter system. The inhibitory effect of radicicol was similar on the signaling of all six steroid hormone receptors and was distinguishable from cytotoxic effects that occurred with higher concentrations. These yeast plasmids provide a high throughput system for comparative assessment of steroid hormone receptor signaling and may be useful in screening for pharmacological or xenobiotic activities.
登录
查看更多内容
影响因子：
8.8
作者：
Leskinen, P;Michelini, E;Virta, M
通讯作者：
Virta, M
用于鉴定亚型选择性雌激素受体配体的二分重组酵母系统
影响因子：
2.6
作者：
Liang, Kaiwei;Yang, Liuqing;Huang, Jian
通讯作者：
Huang, Jian
影响因子：
64.8
作者：
PICARD, D;KHURSHEED, B;YAMAMOTO, KR
通讯作者：
YAMAMOTO, KR
影响因子：
5.3
作者：
Hong, H;Kohli, K;Stallcup, MR
通讯作者：
Stallcup, MR
影响因子：
56.9
作者：
KING, K;DOHLMAN, HG;LEFKOWITZ, RJ
通讯作者：
LEFKOWITZ, RJ