Figure 4: Cis-element binding ability and transcriptional activation assays of McMYB12a and McMYB12b with anthocyanin and PAs biosynthetic genes. | Scientific Reports

Figure 4: Cis-element binding ability and transcriptional activation assays of McMYB12a and McMYB12b with anthocyanin and PAs biosynthetic genes.

From: McMYB12 Transcription Factors Co-regulate Proanthocyanidin and Anthocyanin Biosynthesis in Malus Crabapple

Figure 4

(A) Interaction of McMYB12 proteins with the promoters of flavonoids biosynthetic genes as revealed by yeast one-hybrid assays. The panel shows yeast cells containing distinct effector and reporter constructs grown on an SD/-Trp/-Ura medium plate. Interaction of McMYB12a or McMYB12b, fused to the GAL4 activation domain (pJG4-5-McMYB12a or b), with LacZ driven by the flavonoid biosynthetic gene promoters (pLacZi-promoter of flavonoid biosynthetic gene) is shown in left top panel. Yeast transformed with pJG4-5-McMYB12s/pLacZi, pJG4-5/pLacZi-promoter of flavonoid biosynthetic genes and pJG4-5/pLacZi were used as controls. (B) Gel-shift analysis of McMYB12a binding to the promoters of six flavonoid biosynthetic genes. Purified proteins (10 μg) were incubated with 25 picomoles biotin-labeled probe or mutant probe (mP). For the competition test, non-labeled probes at varying concentrations (from 10- to 1,000-fold excess) or labeled mutant probes (1,000-fold) were added to the above experiment. (C) Gel-shift analysis of McMYB12b binding to the promoters of five flavonoid biosynthetic genes.

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