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Fig. 5. Deletion analysis of FM2 enhancer. (A) Fine mapping of the FM2 enhancer. Blue boxes indicate the domains conserved between human and mouse characterized by sequence identity exceeding 80% over more than 100 bp. Sequences in the boxes denoted by P were used as primers to isolate the counterpart 3.0 kb region in human genome by PCR. Asterisk in HB 0.9 kb indicates residual expression shown in (Be). Percentages on each bar provide the sequence identity of each domain with human or rat. (B) ß-Gal expression driven by the SB 2.8 kb fragment (a), by the BH 1.2 kb fragment (b), by the human 3.0 kb counterpart of the SB 2.8 kb region (c), by the BB 1.0 kb fragment (d), by the HB 0.9 kb fragment (e), by the BB 1.0 kb fragment with mutations in three TCF binding sites (f) and by the BP 200 bp fragment (g) at E10.5 (a-f) and at E11.5 (g). Arrowheads in a and e indicate faint ectopic expression in DRG and residual expression in dorsal midbrain, respectively. Arrows indicate ß-gal expression in cephalic mesenchyme by the 1.8 kb promoter. Scale bars: 400 µm. (C) Alignment of nucleotide sequences of BP 200 bp between mouse and rat.

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