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Fig. 7 | BMC Plant Biology

Fig. 7

From: Transcriptional silencing of 35S driven-transgene is differentially determined depending on promoter methylation heterogeneity at specific cytosines in both plus- and minus-sense strands

Fig. 7

Strategy to assess the direct effect of strand-specific cytosine methylation on promoter activity. a Golden gate cloning to construct the firefly luciferase (Fluc) gene fused with a methylated 35S promoter sequence. Custom-synthesized DNAs containing three methylated cytosine residues (positions − 78, − 66 and − 46) at CG context and three methylated cytosine residues (positions − 84, − 56 and − 33) at CHH context were annealed, digested with BsaI and ligated to the BsaI-digested Fluc gene. b Schematic flow of transient expression assay in protoplasts. In vitro ligation products were directly transfected to N. benthamiana protoplasts, and then luciferase activities in 18 h-incubated protoplasts were measured by fluorometer

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BMC Plant Biology

ISSN: 1471-2229

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