Fig. 3

Viral vector insertion in the def gene caused the def^hi429 mutant phenotype. (A) Schematic drawing shows the genomic DNA structure of the def gene. The viral vector was inserted in the second intron. The positions of two pairs of primers used for genotyping the mutant embryos are indicated, and their PCR products are shown in the DNA gel photos, respectively. Red arrowheads represent the primer pair for checking the viral insertion in heterozygous (+/-) or homozygous (-/-) mutants; green arrowheads for checking the wild-type gene in wild-type sibling (+/+) or heterozygous mutant (+/-). (B) Amino acid sequence alignment of Def (gi37046654) and its homologs/orthologs in human (gi7657019), mouse (gi20340985), Drosophila (gi20130323), C. elegans (gi17555844), and yeast (gi19076047). (C) Injection of def mRNA (def) rescued the def^hi429 mutant phenotype, whereas injection of GFP mRNA (GFP) or mutant def^stop mRNA (data not shown) failed to rescue the mutant phenotype. Embryos 3 and 4 d post-injection were used for whole-mount in situ hybridization with ifabp and trypsin probes, respectively. (mu) Mutant; (wt) wild-type.