Fig 1: Identification of the Pold1 missense mutation in tyrn. (A) Schematic diagrams of the murine Pold1 genomic locus (upper panel) and of the Pold1 domain structure (lower panel). The G2815T nucleotide change (red arrow) was located in exon 23. The corresponding D939Y amino acid substitution was located in the DNA polymerase domain close to the C-terminal domain (CTD); exo: exonuclease domain. (B) Multiple alignments of orthologous Pold1 amino acid sequences around D939Y among different eukaryotic organisms. The mutated aspartate residue was highly conserved. (C) Pold1 expression level in wildtype (left lane) and tyrn (right lane) embryos at E8.5 shown by Western blot. (D) Complementation test crossing strategy (upper panel). Wildtype and tyrn/tm1b embryos were acquired at E7.5 from the complementation test (lower panel). tyrn/tm1b embryos were not able to survive past E7.5. We did not remove the parietal yolk sac from the tyrn/tm1b embryo due to the challenge of dissecting the parietal yolk sac away from the fragile mutant embryo. (E) Crossing strategy to harvest homozygous mutants from heterozygous mice carrying the tyrn allele (upper panel). E7.5 wildtype and mutant embryos (lower panel). n=3 per genotype. Scale bar: 100 µm.
Supplier Page from Abcam for Anti-POLD1 antibody