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Figure 1
Transient expression analysis of the agrp-enhanced, green, fluorescent protein-nitroreductase (agrp-EGFP-NTR) deletion constructs. (A) Schematic diagram of the DNA construct used to generate AgRPNTR (Tg[agrp-EGFP-NTR]) transgenic zebrafish. AgRP (4.7K)-EGFP-NTR contains a 4749 bp 5? flanking region of the agrp gene, and EGFP-NTR fusion has an in-frame 24 bp region downstream of the agrp translational start site. (B) Each deletion construct was generated as described in Section 2.2 for functional analyses of the promoter. A total of 4.6 nL (15 ng/µL) of the deletion constructs for (1) AgRP (2.7K)-EGFP-NTR, (2) AgRP (3.4K)-EGFP-NTR, and (3) AgRP (4.0K)-EGFP-NTR and the full-length construct for (4) AgRP (4.7K)-EGFP-NTR were microinjected into one-cell stage embryos. EGFP signal was observed in 36 h post-fertilization (hpf) embryos injected with these constructs via fluorescence microscopy. Scale bar: 200 µm.