FIGURE 1
Generation of hGNEM743T/WT transgenic lines. (A) A schematic representation of the p(-1.9gne:hGNEM743T/WT; insulin:EGFP) plasmid. Genes, promoters, LTRs, polyadenylation signal and restriction sites are indicated. (B) The hGNE transgenic lines develop normally. The figure presents brightfield images of a WT (top), hGNEWT transgenic (middle), and hGNEM743T transgenic (bottom) larvae at 8 dpf. Scale bar = 1 mm. (C) RT-PCR analysis of hGNE mRNA expression in hGNEM743T and hGNEWT transgenic lines. Agarose gel electrophoresis of RT-PCR amplification products of the hGNE gene (top) and eef1a1a (bottom) as the internal control. Both the hGNEM743T (Mut) and hGNEWT (WT) cDNA samples show the expected amplification in all tested time points and tissues. Non-transgenic WT zebrafish cDNA (WT ZF cDNA) and genomic DNA from transgenic fish (Tg DNA) served as controls for each target. (NTC) no template control, (M) Muscle, (Br) Brain tissue. |