Abstract
The interaction between a protein and DNA is involved in almost all cellular functions, and is vitally important in transcriptional regulation. There are two complementary approaches used to detect the interactions between a transcription factor (TF) and DNA, i.e., the TF-centered or protein–DNA approach, and the gene-centered or DNA–protein approach. The yeast one-hybrid (Y1H) is a powerful and widely used gene-centered system to identify DNA–protein interactions. However, a powerful and simple TF-centered method to study protein–DNA interactions like Y1H is lacking. Here, we provide a TF-centered method based on the Y1H system to identify the motifs recognized by a defined TF, termed TF-centered Y1H. In this system, a random short DNA sequence insertion library is generated as the prey DNA sequences to interact with a defined TF as the bait. TF-centered Y1H could identify quickly the motifs bound by a defined TF, representing a reliable and efficient approach with the advantages of Y1H. Therefore, this TF-centered Y1H may have a wide application in protein–DNA interaction studies.
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Acknowledgments
This work was supported by The National Natural Science Foundation of China (No. 31500535) and The Fundamental Research Funds for the Central Universities (No. 2572017DA01).
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Ji, X., Wang, L., Zang, D., Wang, Y. (2018). Transcription Factor-Centered Yeast One-Hybrid Assay. In: Oñate-Sánchez, L. (eds) Two-Hybrid Systems. Methods in Molecular Biology, vol 1794. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7871-7_11
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DOI: https://doi.org/10.1007/978-1-4939-7871-7_11
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7870-0
Online ISBN: 978-1-4939-7871-7
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