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. 2012 Nov 9;3:247. doi: 10.3389/fpls.2012.00247

FIGURE 2.

FIGURE 2

Schematics of the promoter of sapx used for yeast-one-hybrid screening and binding to the specific cis-element. (A) Fragment psapx1 (–1563 to –1868 bp); psapx2 (–1262 to –1646 bp); psapx3 (–954 to –1321 bp); psapx4 (–609 to –1018 bp); psapx5 (–246 to –691 bp); psapx6 (–1 to –263 bp). Division of psapx2 for localization of the ANAC-binding cis-element: pHis2:sapx2-1/mut (–1432 bis –1646), pHis2:sapx2-2 (–1432 bis –1451). The potential target element for binding of ANAC089 is marked as hatched and the mutagenized region in white box. (B) Experimental identification of the binding site of ANAC089 within promoter segment pHis2:sapx2 using the Y1H system on SD/-Trp/-His/-Leu/+3 AT selective medium (C1, C2, C3) and for control on SD/-Trp/-Leu selective medium (C1-C, C2-C, C3-C). (C1) p His2:sapx2-1; (C2) pHis2:sapx2-1mut; and (C3) p His2:sapx2-2. All cells grew under non-stringent selection showing their viability, while under stringent conditions only the cells grew that were harboring the wild type sapx2-1 fragment and expressing ANAC089.