A bacterial one-hybrid system for determining the DNA-binding specificity of transcription factors

UMMS Affiliation

Program in Gene Function and Expression

Publication Date


Document Type



Animals; Binding Sites; DNA, Bacterial; Drosophila Proteins; Drosophila melanogaster; Escherichia coli; Genes, Regulator; Protein Binding; Protein Interaction Mapping; Sensitivity and Specificity; Sequence Analysis; Transcription Factors; *Two-Hybrid System Techniques


Life Sciences | Medicine and Health Sciences


The DNA-binding specificities of transcription factors can be used to computationally predict cis-regulatory modules (CRMs) that regulate gene expression. 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.

DOI of Published Version



Nat Biotechnol. 2005 Aug;23(8):988-94. Epub 2005 Jul 24. Link to article on publisher's site

Journal/Book/Conference Title

Nature biotechnology

Related Resources

Link to Article in PubMed

PubMed ID