Abstract
DNA-binding proteins, including transcription factors, play essential roles in many biological processes. The identification of the DNA sequences to which these proteins bind is a first, yet still challenging, step for determining their functions. SELEX provides an excellent tool for deciphering protein DNA-binding sequence specificity, and it has been widely adopted for addressing fundamental biological questions (1, 2). SELEX is an experimental procedure that involves the progressive selection, from a large combinatorial double-stranded oligonucleotide library, of DNA ligands with variable DNA-binding affinities and specificities by repeated rounds of partition and amplification. In this chapter, we describe a SELEX protocol that we have successfully applied to both plant and animal MYB transcription factors.
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Acknowledgments
Support in the Grotewold lab for projects involving regulation of gene expression is provided by NRI Grant 2007-35318-17805 from the USDA CSREES, DOE Grant DE-FG02-07ER15881, and NSF grant DBI-0701405. ZX was supported by a 1-year predoctoral Excellence in Plant Molecular Biology & Biotechnology fellowship.
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Chai, C., Xie, Z., Grotewold, E. (2011). SELEX (Systematic Evolution of Ligands by EXponential Enrichment), as a Powerful Tool for Deciphering the Protein–DNA Interaction Space. In: Yuan, L., Perry, S. (eds) Plant Transcription Factors. Methods in Molecular Biology, vol 754. Humana Press. https://doi.org/10.1007/978-1-61779-154-3_14
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DOI: https://doi.org/10.1007/978-1-61779-154-3_14
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