Enhanced yeast one-hybrid assays for high-throughput gene-centered regulatory network mapping.
Enhanced yeast one-hybrid assays for high-throughput gene-centered regulatory network mapping.
复制标题
DOI：
10.1038/nmeth.1748
复制
发表时间：
2011-10-30
期刊：
影响因子：
48
通讯作者：
Walhout, Albertha J. M.
中科院分区：
文献类型：
--
作者：
Reece-Hoyes, John S.;Diallo, Alos;Lajoie, Bryan;Kent, Amanda;Shrestha, Shaleen;Kadreppa, Sreenath;Pesyna, Colin;Dekker, Job;Myers, Chad L.;Walhout, Albertha J. M.
系统生物学的一个主要挑战是了解驱动发育、生理学和病理学的基因调控网络。转录因子和调控基因组区域之间的相互作用提供了基因控制的第一个层次。网关兼容的酵母单杂交（Y1H）检测提供了一种方便的方法来识别和表征的转录因子，可以结合感兴趣的DNA序列的库。然而，为了描绘基因组规模的调控网络，需要以高通量和高覆盖率处理大量DNA片段。在这里，我们提出了“增强型”Y1H（eY1H）检测，该检测利用机器人交配平台，配备一套改进的Y1H试剂和自动读数定量。我们证明，eY1H检测提供了良好的覆盖范围，并确定在短时间内多个DNA片段的相互作用的转录因子。eY1H检测将成为秀丽隐杆线虫和其他模式生物以及人类基因调控网络作图的重要工具。
A major challenge in systems biology is to understand the gene regulatory networks that drive development, physiology and pathology. Interactions between transcription factors and regulatory genomic regions provide the first level of gene control. Gateway-compatible yeast one-hybrid (Y1H) assays present a convenient method to identify and characterize the repertoire of transcription factors that can bind a DNA sequence of interest. To delineate genome-scale regulatory networks, however, large sets of DNA fragments need to be processed at high throughput and high coverage. Here, we present “enhanced” Y1H (eY1H) assays that utilize a robotic mating platform with a set of improved Y1H reagents and automated readout quantification. We demonstrate that eY1H assays provide excellent coverage and identify interacting transcription factors for multiple DNA fragments in a short amount of time. eY1H assays will be an important tool for gene regulatory network mapping in Caenorhabditis elegans and other model organisms, as well as humans.
登录
查看更多内容
影响因子：
10.5
作者：
Lesch, Bluma J.;Gehrke, Andrew R.;Bargmann, Cornelia I.
通讯作者：
Bargmann, Cornelia I.
影响因子：
9.9
作者：
通讯作者：
--
影响因子：
4.5
作者：
Tabuchi, Tomoko M.;Deplancke, Bart;Hagstrom, Kirsten A.
通讯作者：
Hagstrom, Kirsten A.
影响因子：
7
作者：
Deplancke, B;Dupuy, D;Walhout, AJM
通讯作者：
Walhout, AJM
影响因子：
4.8
作者：
Walhout, AJM;Vidal, M
通讯作者：
Vidal, M