title: A bacterial one-hybrid system for determining the DNA-binding specificity of transcription factors.
abstract: 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.
authors.affiliation: Program in Gene Function and Expression, University of Massachusetts Medical School, 364 Plantation St., Worcester, Massachusetts 01605, USA.
authors.forename: Xiangdong
authors.initials: X
authors.lastname: Meng
authors.forename: Michael H
authors.initials: MH
authors.lastname: Brodsky
authors.forename: Scot A
authors.initials: SA
authors.lastname: Wolfe
mesh_terms: D000818:Animals; D001665:Binding Sites; D004269:DNA, Bacterial / Q000737:chemistry* / Q000378:metabolism*; D029721:Drosophila Proteins / Q000737:chemistry / Q000378:metabolism*; D004331:Drosophila melanogaster; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005809:Genes, Regulator; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D012680:Sensitivity and Specificity; D017421:Sequence Analysis / Q000379:methods*; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
chemical_list: D004269:DNA, Bacterial; D029721:Drosophila Proteins; D014157:Transcription Factors