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. 2009 Oct 9;4(10):e7354. doi: 10.1371/journal.pone.0007354

Figure 2. Detection of stop codon readthrough in stably transformed Arabidopsis plants.

Figure 2

(A) Reporter gene constructs introduced into the Arabidopsis nuclear genome. The strong constitutive p045 promoter (open arrow) was used to drive expression of NAN-GUS dual reporter constructs containing either the wild-type TMV leaky stop codon region (p045:TMV-TAG) or the NRStop region (p045:TMV-NRStop) in stably transformed plants. (B) Histochemical analysis of progeny of single insertion transgenic Arabidopsis lines. Each panel contains six seedlings, 3 stained for NAN activity (left half) and 3 stained for GUS activity (right half). NAN activity stains blue, GUS activity magenta. Seedlings expressing a p045:GUS control construct show, as expected, strong GUS activity but no NAN activity (top panel). Seedlings expressing the p045:TMV-NRStop construct show strong NAN activity but negligible GUS activity (bottom panel), while seedlings expressing p045:TMV-TAG show strong NAN activity and low, but significant GUS activity (middle panel). (C) GUS/NAN activity ratios of seedlings of three individual transgenic lines expressing the p045:TMV-TAG (lines 7, 10, 11) or the p045:TMV-NRStop (lines 8, 11, 12) construct. NAN and GUS activity was quantified in total protein extracts from two pools of twelve seedlings of each line.