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Fig. 3. Mapping of regulatory motifs essential for PLE-specific activity of the
FoxE3 enhancer. (A-C) Representative transgenic embryos
(stages 22-24) generated with the GFP reporter constructs shown on the left.
Black arrows indicate the PLE. Numbers of embryos with GFP expression in the
PLE and the total number of normally (or near normally) developing embryos
injected with each construct are indicated on the right-hand side with
percentages of the GFP-positive cases. The white line in A indicates the plane
of the transverse section shown in the inset. The black arrow in the inset
indicates GFP expression in the PLE overlying the optic vesicle. The embryo
shown in C was generated by co-transgenesis, i.e. co-injection of the 462 bp
enhancer of Xenopus FoxE3 amplified by PCR along with the βGFP
cassette. (D) Identification of transcription factor-binding motifs
essential for PLE-specific expression by mutation analysis. wt is the
construct used in Fig. 3A (Xt462-βGFP). mt1-mt9 were generated from
wt/Xt462-βGFP by introducing a base-substitution mutation (cross) into
each of the conserved transcription factor-binding motifs. The bar chart shows
the percentage of the embryos that showed GFP expression in the PLE among
total developed embryos injected with the constructs shown on the left. Actual
numbers of GFP-positive cases and total numbers of scored embryos are
indicated in parentheses.
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