MMAPPR results for zy14. (A,B) Genome-wide (A) and chromosome 5 (B) Loess fit results from mapping the zy14 line. (•) Centromere location; (* in B) location of the mutation. (C,D) Sanger sequencing traces from a wild-type sibling and a zy14 mutant, respectively. (Arrow) A single base changed in the mutant; (* and underline) the resulting stop codon. (E–H) Fluorescent images of the vascular phenotype visualized using the Tg(kdrl-EGFP) line at 40 hpf, which marks vasculature. (E) Wild-type zy14 sibling. (F) zy14 mutant. (G) zy14 mutant rescued by injection of wild-type cds2 mRNA. (H) Failure of cds2 mutant mRNA injection to rescue zy14 mutant embryo. In three experiments, uninjected embryos from heterozygote crosses had wild-type phenotypes in 39/51, whereas 109/109 embryos injected with wild-type cds2 mRNA had a wild-type phenotype, indicative of rescue. In contrast, mutant cds2 mRNA with the single-base change seen in zy14 was not able to rescue: 18/26 injected embryos had wild-type phenotype, compared with 14/20 uninjected siblings.
