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Figure 2


Fig. 2. The eve locus contains a Pho-dependent PRE. (A,B) Eyes of adult flies closely matched in age (based on body color) and within 14 hours of eclosion (during which time no discernable changes in eye color occurred), either heterozygous (A) or homozygous (B) for transgenic PRE300 in a white mutant background. Note the dramatically lighter color in B, owing to PSS of transgenic mini-white. (C,D) Eyes of pharate adults carrying a homozygous PRE300 transgene in a heterozygote (C; indistinguishable from wild-type eyes in this line) or homozygous (D) pho1 mutant background. Note the slightly darker eye color in D compared with C. (E,F) Eyes of pharate adults carrying a homozygous GFP-RR transgene in a heterozygous (E; indistinguishable from wild-type eyes in this line) or homozygous (F) pho1 mutant background. GFP-RR (described in the Materials and methods) carries the neuronal RP2+a/pCC enhancer and PRE300 (+7.9 to +9.2 kb) (Fujioka et al., 2003). As with PRE300 alone (Fig. 1), GFP-RR transgenic lines showed PSS (64%: 7 out of 11 lines). Note the darker eye color in F. (G-L) Anti-β-gal-stained stage 13 embryos carrying a bxd-Ubx-Z-derived transgene. (G-I) Transgenes (indicated below) inserted at attP-target site 95E5. (G) {lambda}500-bxd-Ubx-Z, a non-PRE-containing negative control. (H) PRE300-bxd-Ubx-Z. Note the lack of β-gal expression in the anterior region indicated by the black line, relative to G. (I) PRE300{Delta}Pho-bxd-Ubx-Z. Note that mutation of the Pho-binding site caused de-repression of β-gal in many cells in the anterior regions, indicated by arrowheads (compare with H). (J-L) PRE300-bxd-Ubx-Z inserted at attP-target site 52D (Bateman et al., 2006). (J) Wild-type background. (K) Pc4 background. (L) ph503 background. Note de-repression of β-gal in the anterior region indicated by the black line in K and L, relative to J. (M-O) Anti-β-gal-stained CNS from stage 15 embryos carrying transgenes (indicated below) inserted at attP-target site 95E5. (M) {lambda}500-bxd-Ubx-Z. (N) PRE300-bxd-Ubx-Z. Note the lack of β-gal expression in the anterior region indicated by the black bracket, relative to M. (O) PRE300{Delta}Pho-bxd-Ubx-Z. Note that mutation of the Pho-binding site caused de-repression of β-gal in many cells in the anterior region indicated by the black bracket, relative to N.





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