Fig. 5

The samba^hkz2 mutation is mapped to the haus3 gene in linkage group 7 of the zebrafish genome. (A) Positional cloning information. The samba^hkz2 mutation is mapped to a three-overlapping-BAC region between markers CR318672_119281 and CR318603_55485 in LG7. This region contains eight candidate genes (blue). The simple sequence length polymorphism markers (SSLP) or single nucleotide polymorphism (SNP) markers used in mapping are labeled in red with their respective recombinants number (out of a total of 1910 homozygous samba^hkz2 embryos) in green. (B) Sequencing results showing a nonsense mutation (a T-to-G transition) in haus3 (zgc:153228) gene. (C) Gene structure of haus3 (zgc:153228). Gap region indicates the absence of 48 bp in haus3-α (zgc:153228-α). (D-F) Immunohistochemical staining with the anti-V5 monoclonal antibody on pcDNA3.1/nV5-haus3-β-transfected cos-7 cells and DAPI staining of the same transfected cells. (G-J) In vitro synthesized full-length haus3 RNA rescues the mutant phenotype. WISH of cmyb expression in wild type (G), uninjected samba^hkz2-/- mutant control (H) and samba^hkz2-/- mutant embryos injected with either wild-type haus3-α (I) or haus3-β RNA (J). The phenotype seen in I and J is representative of 10 out of 38 and 18 out of 55 mutant embryos observed, as indicated.