The zebrafish mutant gavi. At 48 hpf, staining for hemoglobin with o-dianisidine revealed that the homozygous gavi (gav) mutant embryo (A) has decreased hemoglobin staining compared with a wild-type sibling (B). The scale bar represents 200 μm. Intermediate resolution mapping of the gavi mutation, using bulk segregant analysis, revealed that the mutation was on linkage group 2, near the transferrin-a locus (C). The green box represents the centromere. A protein sequence alignment (D) illustrates the relationship of the zebrafish transferrin-a protein sequence to transferrins from other species. Conserved residues are shaded light blue. The predicted truncation sites in the zebrafish mutants are indicated by asterisks: gav^IT029 (red) and gav^HE067 (blue).