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. 2007 Jul 10;35(14):4882–4894. doi: 10.1093/nar/gkm519

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© 2007 The Author(s)

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — Verification of the system for monitoring the functional interactions between the upstream and the core promoter sequences. In each panel, the schematic on the left indicates the reporter plasmids introduced into fertilized eggs. (A) Expression of CFP/YFP reporter genes driven by the same promoters. Three sets of CFP/YFP reporter plasmids were used: pM4705/pM4692, driven by the wild-type HpOtxE promoter; pM4725/pM4731, driven by the wild-type SpSpec2a promoter and pM4694/pM4706, driven by a mutated HpOtxE promoter bearing the TATAAA sequence instead of the TATTCA sequence (OtxE^M; changed nucleotides are shown in red). The expression of these reporter genes were examined with fluorescence microscopy at the blastula, early gastrula, mid gastrula, late gastrula and pluteus stages, as indicated at the top. The number of embryos showing the expression pattern corresponding to each profile (defined in Figure 2) is summarized in the right panel. The vertical and horizontal axes indicate the number of embryos and expression pattern profiles, respectively. (B) Expression of CFP/YFP reporter genes driven by promoters containing or not containing the upstream sequence. (Upper panel) The reporter plasmids used were pM4705/pM4915 (I) or pM4914/pM4692 (II), which are driven by the wild-type HpOtxE promoter and its core promoter connected to CFP and YFP, respectively (I), or vice versa (II). The expression of the reporter genes was examined with fluorescence microscopy at different developmental stages, as indicated at the top. The number of embryos showing the expression patterns corresponding to each profile is summarized in the right panel as described in A. The color of the bars representing profile #4 reflect the signal that was observed (CFP, green; YFP, red). (Lower panel) Two CFP/YFP reporter plasmids were used, pM4725/pM4917 (I) and pM4916/pM4731 (II), which are driven by the wild-type SpSpec2a core promoter fused or not fused with the HpOtxE upstream sequence and connected to the complementary patterns of CFP and YFP. The number of embryos showing the expression patterns corresponding to each profile is summarized in the right panel as described above.