Novel cell surface luciferase reporter for high-throughput yeast one-hybrid screens.

Novel cell surface luciferase reporter for high-throughput yeast one-hybrid screens.
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DOI:
10.1093/nar/gkx682
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发表时间:
2017-10-13
影响因子:
14.9
通讯作者:
Pruneda-Paz JL
Pruneda-Paz JL
中科院分区:
生物学2区
文献类型:
--
作者:
Bonaldi K;Li Z;Kang SE;Breton G;Pruneda-Paz JL

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以基因为中心的酵母单杂交(Y1H)筛选为鉴定转录因子(TF)与启动子的相互作用提供了一种强大而有效的策略。虽然全基因组的Tf或Feome克隆收集越来越多,但筛选方案对用于识别相互作用的酶反应的性质和以高通量格式执行分析所需的程序有固有的限制。在这里,我们提出了一种简化的策略的开发和验证,用于定量和全自动化的以基因为中心的Y1H筛查,使用一种新的细胞表面Gaussia荧光素酶报告。
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.
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