Fig. 2

Fig. 2 Transgenic zebrafish line Tg(LCR^RH2-RH2-1:GFP)^pt112, expresses GFP but not GST-His fusion in the retina. (A) The schematic illustrates the structures and tandem arrangement of the GST-His and GFP reporter genes in the transgene construct that was used to generate the Tg(LCR^RH2-RH2-1:GFP)^pt112 line. The regions amplified by PCR genotyping reactions are indicated with double-headed arrows. (B) In the adult Tg(LCR^RH2-RH2-1:GFP)^pt112 retina at 8 mpf (months postfertilization), the expression of the GST-His protein was undetectable, whereas GFP expression is very strong. (C) RT-PCR and genomic DNA PCR analyses demonstrated that the GST-His gene was present in the Tg(LCR^RH2-RH2-1:GFP)^pt112 genome, and was apparently transcribed at both 5 dpf and 9 mpf. (D) Western blotting analyses confirmed that GST-His fusion protein was not expressed in Tg(LCR^RH2-RH2-1:GFP)^pt112 fish at both 5 dpf and 9 mpf. The asterisks indicate the position where the fusion protein was supposed to be. Unrelated His-Tagged and GST-tagged recombinant proteins expressed in Escherichia coli were used as controls for Western blotting conditions.