Fig. 1

The tortuga^b644 allele is a 1.46 Mb deficiency that includes the pnrc2 gene. Haploid-based mapping revealed that the tortuga lesion lies 0.30 cM to the left (4/1434 recombinants) and 0.37 cM to the right (4/1083 recombinants) of the SSLP markers z13511 and z9511, respectively (A). The extent of the deletion was refined by PCR-based screening of regions within and around the deletion interval from pooled genomic DNA samples of 10 wild-type (WT) and 10 tortuga mutant embryos (B); regions that fail to amplify in mutants are deleted in the tor^b644 allele (A, B). Among BACs spanning the deletion interval, only BAC AL844887 restores her1 expression when injected into tor mutants (A; Fig S1A-D). In wild-type embryos, her1 is expressed in a striped pattern (n=54/54) (C). In contrast, embryos injected with 6 ng of pnrc2 splice-blocking morpholino (sbMO) have a tortuga-like her1 expression defect (n=37/39) (D). When pnrc2 mRNA is co-injected with 4 ng pnrc2 sbMO, her1 expression is partially restored in a dose-dependent manner (n=3/19 WT her1 expression, 150 pg pnrc2 mRNA; n=7/17 WT her1 expression, 600 pg pnrc2 mRNA) (E; Table 1). Alignment of vertebrate Pnrc2 amino acid sequences reveals a conserved 10 amino acid N-terminus (purple) and conserved C-terminal SRC-Homology 3 (SH3) (red) and Nuclear Receptor (NR box) domains (green) (F). Clustal alignments were performed in consultation with published alignments for vertebrate Pnrc2 (Valkov et al., 2016).