Nature Biotechnology
23, 988 - 994 (2005)
Published online: 24 July 2005; | doi:10.1038/nbt1120
A bacterial one-hybrid system for determining the DNA-binding specificity of transcription factorsXiangdong Meng1, Michael H Brodsky1, 3
& Scot A Wolfe1, 21
Program in Gene Function and Expression, University of Massachusetts Medical School, 364 Plantation St., Worcester, Massachusetts 01605, USA. 2
Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 364 Plantation St., Worcester, Massachusetts 01605, USA. 3
Program in Molecular Medicine, University of Massachusetts Medical School, 364 Plantation St., Worcester, Massachusetts 01605, USA.
Correspondence should be addressed to Scot A Wolfe scot.wolfe@umassmed.edu The DNA-binding specificities of transcription factors can be used to computationally predict cis-regulatory modules (CRMs) that regulate gene expression1. However, the absence of specificity data for the majority of transcription factors limits the widespread implementation of this approach. We have developed a bacterial one-hybrid system that provides a simple and rapid method to determine the DNA-binding specificity of a transcription factor. Using this technology, we successfully determined the DNA-binding specificity of seven previously characterized transcription factors and one novel transcription factor, the Drosophila melanogaster factor Odd-skipped. Regulatory targets of Odd-skipped were successfully predicted using this information, demonstrating that the data produced by the bacterial one-hybrid system are relevant to in vivo function.
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