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• W2805422023 abstract "Identification of transcription factor (TF)–promoter interactions is key to understanding the basic molecular underpinnings of gene regulation. The complexity of gene regulation, however, is driven by the combined function of several TFs recruited to the promoter region, which often confounds the discovery of transcriptional regulatory mechanisms. Genome sequencing enabled the construction of TF-specific ORFeome clone collections that can be used to study TF function with unprecedented coverage. Among the recently developed methods, gene-centered yeast one-hybrid (Y1H) screens performed with these ORFeome collections provide a simple and reliable strategy to identify TF–promoter interactions. Here, we describe high-throughput cloning protocols used to generate a gold standard TF ORFeome collection for the model organism Arabidopsis thaliana. Furthermore, we outline the protocol to build a daughter clone collection suitable for the Y1H assay and a high-throughput Y1H screening procedure that enables rapid assessment of thousands TF–promoter interactions using a robotic platform. These protocols can be universally adopted to build ORFeome libraries and thus expand the usage of gene-centered Y1H screens or other alternative strategies for discovery and characterization of TF functions." @default.
• W2805422023 created "2018-06-13" @default.
• W2805422023 creator A5033883138 @default.
• W2805422023 creator A5038875814 @default.
• W2805422023 creator A5074324847 @default.
• W2805422023 date "2018-01-01" @default.
• W2805422023 modified "2023-10-16" @default.
• W2805422023 title "Construction of Arabidopsis Transcription Factor ORFeome Collections and Identification of Protein–DNA Interactions by High-Throughput Yeast One-Hybrid Screens" @default.
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• W2805422023 doi "https://doi.org/10.1007/978-1-4939-7871-7_10" @default.
• W2805422023 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/29855956" @default.
• W2805422023 hasPublicationYear "2018" @default.
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