Fig. 4

bonaparte and poppy are mutant alleles of basonuclin-2 (bnc2).

(A) Mapping of bonaparte to chromosome 1. (B,C) Identification of premature stop codons in bnc2 alleles of bonaparte (B) and poppy (C). Shown are wild-type AB^wp alleles, homozygous mutant alleles, and heterozygous mutant alleles. *, stop codon. (D) Phylogenetic analysis places zebrafish bnc2 within the clade of other vertebrate bnc2 loci. Values in at nodes are bootstrap support based on 1,000 replicates (PAUP heuristic parsimony analysis) followed by posterior probabilities from Bayesian analysis. Abbreviations: Dr, Danio rerio; Ol, Oryzias latipes; Ga, Gasterosteus aculeatus; Tr, Takifugu rubripes; Xt, Xenopus tropicalis; Gg, Gallus gallus; Md, Monodelphis somestrica; Cf, Canis familiaris; Mm, Mus musculus; Rn, Rattus norvegicus; Mam, Macaca mulatta; Pt; Pan troglodytes; Hs, Homo sapiens; Aa, Aedes aegypti; Ag, Anopheles gambiae. (E) Domain structure of bnc2 and predicted truncated proteins. (F) Northern blot of poly(A)+ mRNA extracted from whole fish at the times indicated and probed with a full length bnc2 RNA probe. Two major bnc2 isoforms were detected. Equal loading verified by Nanodrop spectrophotometry and β-actin control probe (not shown). (G) bnc2-GFP fusion protein shows nuclear localization in embryo epidermis at 24 hours post-fertilization.