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

被引:11
作者
Bonaldi, Katia [1 ,2 ]
Li, Zheng [1 ,2 ]
Kang, S. Earl [1 ,4 ]
Breton, Ghislain [3 ]
Pruneda-Paz, Jose L. [1 ,2 ]
机构
[1] Univ Calif San Diego, Div Biol Sci, La Jolla, CA 92093 USA
[2] Univ Calif San Diego, Ctr Circadian Biol, La Jolla, CA 92093 USA
[3] McGovern Med Sch, Dept Integrat Biol & Pharmacol, Houston, TX 77030 USA
[4] Univ Georgia, Dept Plant Biol, Athens, GA 30602 USA
基金
美国国家科学基金会; 美国国家卫生研究院;
关键词
GENE REGULATORY NETWORK; TRANSCRIPTION FACTORS; CIRCADIAN CLOCK; SACCHAROMYCES-CEREVISIAE; FUNCTIONAL GENOMICS; BETA-GALACTOSIDASE; ARABIDOPSIS; ASSAYS; EXPRESSION; SYSTEM;
D O I
10.1093/nar/gkx682
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页数:12
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