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Fig. 2
The melanophore integrity gene maps to a 53 kilobase region on chromosome 10.
A) Flanking CA repeat markers and their respective number of recombinants per meioses (shown in red and blue) narrowed the region down to 53 kilobases, containing the ift46, vps11 and hyou1 genes. Being the most likely candidate (see text for details), vps11 cDNA was generated and sequenced. B) Sequenced vps11 cDNA indicated a single base pair deletion at base pair 37 which causes a frameshift and early stop at base pairs 5355. Other differences in sequenced vps11 cDNA include a single base pair substitution at base pairs 615 and 2183. C) RT-PCR on wildtype and integrity mutant mRNA indicates that vps11 transcripts are present in integrity mutants. vps11 cDNA was amplified in regions “a”, “b” and “c” as indicated in 2B (blue arrows). D) Immunoblot and densitometry analysis of Vps11 protein expression in wildtype (AB) and vps11w66 mutant larvae. Vps11 protein is not expressed or is expressed at sub detection levels. E-H) Dorsal head images of live, 6dpf zebrafish larvae born from heterozygous vps11 and platinum parents (E), heterozygous vps11 parents (F and H) and heterozygous platinum parents (G). We observed the integrity phenotype in ~25% of the clutch resulting from heterozygous vps11 and platinum matings.