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Fig. 5. Identification of the promoter-tethering element. On the B-A-5
construct (top diagram), the IAB5 enhancer specifically bypasses the
abd-A promoter (B) to activate expression from the
Abd-B promoter (A). When the 255 bp region extending from -40
to -294 bp relative to the transcription start site is removed from the
Abd-B promoter (B
PTE-A-5 construct; middle
diagram), the IAB5 enhancer is now re-directed to the abd-A promoter,
as no CAT expression is detected (C) and strong lacZ
expression is detected (D) in the majority of embryos. Extremely weak
CAT expression in an IAB5-directed pattern was detected in a few
transgenic embryos (data not shown). The integrity of the
Abd-B
PTE promoter was confirmed
by insertion of the IAB2 enhancer 3' of the
Abd-B
PTE-CAT reporter gene on
the 2-B
PTE-A-5 construct (bottom diagram). This resulted in
IAB2 activation of the Abd-B
PTE
promoter indicated by an IAB2-driven CAT expression pattern in
parasegments 7 (black arrowhead), 9, 11 and 13 in transgenic embryos
(E), whereas lacZ is activated in an IAB5-driven expression
pattern in the more posterior parasegments 10, 12 and 14 (F). Embryos
exhibiting weaker staining are shown to clearly demonstrate the different
expression patterns driven by the two IAB enhancers, although overall the
expression in the 2-B
PTE-A-5 embryos was comparable to other
transgenic lines analyzed in this study.