A bacterial one-hybrid system for determining the DNA-binding specificity of transcription factors.
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.
Meng
Xiangdong
X
Program in Gene Function and Expression, University of Massachusetts Medical School, 364 Plantation St., Worcester, Massachusetts 01605, USA.
Brodsky
Michael H
MH
Wolfe
Scot A
SA
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
D004269:DNA, Bacterial; D029721:Drosophila Proteins; D014157:Transcription Factors