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Fig. 1 Zebrafish Tgfbr3 gene map, exon 1 assignment and promoter sequence and activity. (A) The 16 Tgfbr3 exons are shown as black numbered rectangles on the line representing of the 300 000 bp in Danio rerio chromosome 6 containing the Tgfbr3 gene. The location of the testis-specific bromodomain gene (BRDT), transcribed from the opposite strand, is indicated. The position of the bacterial artificial chromosome (BAC) DKEY-197A14 and its zoomed-in ApaLI-ApaLI fragment containing the BG promoter are shown. Positions of primers 3.20, 3.13, and 3.15, located in exons 1, 5, and 9, respectively, are indicated by arrowheads. (B) Zebrafish Tgfbr3 promoter nucleotide sequence. The nucleotide sequence of the ApaLI-ApaLI fragment containing the Tgfbr3 promoter and first exon is shown. Positive ascending numbers on the right are from upstream ApaLI to downstream ApaLI sites. Negative descending numbers on the left are upstream from the sequences assigned as exon 1 (bold italics); nucleotides belonging to following intron 1 are shown in lower case letters. Position of primer 3.20 is indicated by dotted underline. Relevant responsive elements identified by Mat Inspector are underlined and named. Position of relevant restriction sites are indicated with their names above the sequence. (C) The RT-PCR products obtained with primers 3.20 to 3.13 (lane 1) and primers 3.20 to 3.15 (lane 2) are separated, along with 100-bp molecular markers ladder (lane M), by electrophoresis in an agarose gel. (D) The graph shows the beta-galactosidase-normalized luciferase activity obtained upon transfection of ZF4 fibroblasts with the promoter-less pGL3-basic plasmid or its derivate, p3.2tgfbr3:luc, which contains the ApaLI-BsaJI 3260 bp fragment.