A yeast one-hybrid assay tests candidate protein–DNA interactions by connecting them to reporter gene expression in yeast. In a common design, a chosen DNA sequence is placed near a reporter gene; a candidate protein is joined to a transcriptional activation domain. Binding that recruits the activation domain can turn on the reporter. The measured response is thus a genetically constructed readout of a possible interaction.[ref]
Matchmaker Gold Yeast One-Hybrid Library Screening System User Manual by Clontech Laboratories. pp.4–6: bait, prey and reporter principles
For example, compare the same candidate protein with an intact candidate binding sequence and a changed sequence, together with a negative control lacking the candidate. Reporter activation restricted to the intact sequence supports sequence-dependent recruitment. Reporter activation without the candidate instead reveals background activation that can mimic an interaction.[ref]
Matchmaker Gold Yeast One-Hybrid Library Screening System User Manual by Clontech Laboratories. p.24: genuine and false-positive reporter activation
The assay helped identify auxin response factor 1, which binds an auxin response element. Such a positive result is evidence about binding in the yeast reporter context. Establishing control of the corresponding gene in a plant still requires evidence in that biological context; binding and a physiological gene expression response are separate questions.[ref]
ARF1, a transcription factor that binds to auxin response elements by T. Ulmasov and others. Abstract: factor isolation and comparison of binding specificity with auxin responsiveness
Articles by others on the same topic
There are currently no matching articles.