Bonaldi Katia, Li Zheng, Kang S Earl, Breton Ghislain, Pruneda-Paz Jose L
Division of Biological Sciences, University of California San Diego, La Jolla, CA 92093, USA.
Center for Circadian Biology, University of California San Diego, La Jolla, CA 92093, USA.
Nucleic Acids Res. 2017 Oct 13;45(18):e157. doi: 10.1093/nar/gkx682.
Gene-centered yeast one-hybrid (Y1H) screens provide a powerful and effective strategy to identify transcription factor (TF)-promoter interactions. While genome-wide TF ORFeome clone collections are increasingly available, screening protocols have limitations inherent to the properties of the enzymatic reaction used to identify interactions and to the procedure required to perform the assay in a high-throughput format. Here, we present the development and validation of a streamlined strategy for quantitative and fully automated gene-centered Y1H screens using a novel cell surface Gaussia luciferase reporter.
以基因为中心的酵母单杂交（Y1H）筛选提供了一种强大而有效的策略来鉴定转录因子（TF）与启动子的相互作用。虽然全基因组TF开放阅读框克隆文库越来越容易获得，但筛选方案存在一些局限性，这些局限性源于用于鉴定相互作用的酶促反应的特性以及以高通量形式进行检测所需的程序。在这里，我们展示了一种简化策略的开发和验证，该策略使用新型细胞表面高斯荧光素酶报告基因进行定量且完全自动化的以基因为中心的Y1H筛选。
