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. 2014 Feb 13;15(2):2517–2537. doi: 10.3390/ijms15022517

Figure 5.

Figure 5.

Promoter construct and co-transfection assays. (A) Partial sequence of the MBSII-35Smin-GUS construct. The tandem MBSII element is highlighted; the minimal CaMV35S promoter region is underlined. The GUS start codon is shown in bold. For comparison, the DNA fragment efficiently bound by MYB44 [40] is shown in the top line; and (B) Promoter/effector relations were studied in Arabidopsis protoplasts. Arabidopsis mesophyll protoplasts were co-transfected with the reporter constructs MBSII::GUS or Rd29A::GUS, plus one of the following effector constructs: YFP (control), MYB44 or MYB44-REP. β-glucuronidase activity was assessed in protein extracts by quantitative MUG assay. Conversion of the substrate was measured 4 h after incubation at 37 °C. Six aliquots of protoplasts were transformed for each promoter/effector combination. Protoplast transformation of 35S::GUS for constitutive expression of the reporter gene served as positive control. Note that GUS expression from the 35S promoter is approximately 100 times higher than in MBSII or Rd29A. The experiment was repeated twice with similar results.