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Fig. 7. Effects of the SAC51 5'-leader on GUS
reporter gene expression. (A) The GUS gene constructs
consisting of the SAC51 promoter with either wild-type SAC51
or the mutant sac51-d 5'-leader sequences and the GUS
coding sequence. White arrows represent the SAC51 promoter, boxes
indicate exons, lines indicate introns; gray boxes represent uORFs and black
blocks correspond to the GUS-coding sequence. (B-F) GUS
staining (blue) in transgenic plants harboring the SAC51-GUS
construct (left) or the sac51-d-GUS construct (right).
Sixteen-day-old seedlings (B), inflorescences (C), young rosette leaves (D),
roots (E), and mature embryos (F) are shown. Scale bars: B,C, 5 mm; D,E, 1 mm;
F, 100 µm. (G) The relative GUS translational efficiency of
SAC51-GUS and sac51-d-GUS constructs in wild-type (WT),
acl5-1 and sac51-d acl5-1 backgrounds, and that of the
sac51-C549A-GUS (C549A) construct in the wild-type background. The
levels of GUS activity and GUS mRNA in the SAC51-GUS
transgenic line in the wild-type background were set at 1.0. The GUS
translational efficiencies were calculated as the GUS activity divided by the
GUS mRNA level for each plant. The GUS mRNA levels were
normalized to the ACT8 level in each sample. Bars show
mean±s.d. (n=3). (H) Nucleotide and amino acid changes
at the sac51-d mutation site in SAC51-GUS, sac51-d-GUS and
sac51-C549A-GUS constructs. Nucleotide and amino acid changes are in
bold.
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