Fig. 3

Fig. 3 Detection of a nonsense point mutation in gata1. (a) The chromatograms show sequence of PCR products derived from a homozygous wild-type embryo (+/+) (Upper) and a homozygous mutant embryo (-/-) (Lower) for the gata1 gene nt 1013-1017. (b) Genotyping of vlt^m651 embryos. PCR products using primers Arg-339-S and Arg-339-AS synthesized from embryo DNA of vlt^m651 incrosses were digested with TaqI and electrophoresed on a 2% agarose gel. After TaqI digestion, mutant alleles appear as 246-bp and wild-type alleles as 121- and 125-bp products, which migrate together in the gel shown. Phenotypically wild-type embryos (WT) identified by presence of circulating blood and phenotypically mutant (M) embryos identified by absence of circulating blood are shown. (c) A schematic representation of the N-finger, C-finger, and basic domain of Gata1 and a sequence alignment of the basic domain from different species zebrafish (zf), human (h), mouse (m), and Xenopus (x) are shown with the Arg-339 → Stop mutation identified by a STOP. The asterisks mark the aminoacids that make direct contact with the minor groove of DNA (29).