= Yeast one-hybrid assay
A \b[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.<Matchmaker Gold Yeast One-Hybrid Library Screening System User Manual by Clontech Laboratories>[pp.4–6: bait, prey and reporter principles]{ref}
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.<Matchmaker Gold Yeast One-Hybrid Library Screening System User Manual by Clontech Laboratories>[p.24: genuine and false-positive reporter activation]{ref}
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.<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]{ref}
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