Figure 1

Genetic identification of rpgrip1 mutant zebrafish. (A) Schematic structure of human RPGRIP1, zebrafish Rrpgrip1 (zRPGRIP1) and truncated zebrafish rpgrip1 mutant. (B) Sequence validation of the C to T conversion in homozygous zebrafish, which resulted in a nonsense mutation and changing glutamine codon into stop codon (CAG to TAG, indicated with a box). (C) Gel picture of BbvI digested PCR products amplified using wild type and heterozygous fish genomic DNAs as templates. The PCR fragment was 99 bp and BbvI cut the fragment amplified from wildtype zebrafish to 59 bp and 40 bp, but could not cut the fragment amplified from mutant zebrafish. Wildtype zebrafish (rpgrip1 ^+/+) showed only one band because both bands (59 bp and 40 bp) are too close and could not be well separated. The heterozygous zebrafish, rpgrip1 ^+/−, showed two bands: the upper band is for the mutant PCR product, the lower band is BbvI-cut PCR product (59 bp and 40 bp). The homozygous mutant zebrafish, rpgrip1 ^−/−, showed one band, the 99 bp fragment, which BbvI could not cut. UD, undigested PCR product; D, BbvI-digested fragments.