Figure 2
Dnahc5 mutation involves an in-frame DNA deletion.
(A) Genomic DNA amplification using primers situated in introns 6 and 17 of the Dnahc5 gene yielded a 3.3-kb DNA fragment from a mutant embryo exhibiting heterotaxy (lane 4), while no product was obtained in DNA from normal control embryos (lanes 2, 3, and 5) or no template control (lane 1). (B) Dnahc5 cDNA sequencing showed that exon 6 is contiguous with sequence in exon 18 (arrow), indicating deletion of exons 7–17. (C) The upper bar depicts the exon/intron organization of the mouse Dnahc5 gene. Immediately below is a schematic of the Dnahc5 mRNA transcript. The red box delineates the region deleted in the Dnahc5del593 mutant. The Pfam domains in the protein (http://pfam.sanger.ac.uk/) are shown, including dynein heavy chain N-terminal domains 1 and 2 (DHC_N1 and _N2; residues 246–804 and 1,397–1,809, respectively) and dynein heavy chain domain (residues 3,924–4,619). Two Pfam ATPase domains (residues 2,254–2,398 and 2,582–2,729) were not shown for clarity. (D) Expanded view showing region of the Dnahc5 gene containing DNA insertion derived from other chromosomes. Red: 401 bp of 31-bp tandem repeat region 99% identical to Chr4:131,017,719–131,018,166 (excluding a 47-bp gap); yellow: 265 bp of the 3′-untranslated region of the Csnk2a1 gene (Chr2:151,972,987–151,973,251); green: 1,204 bp of the last exon of the Zbtb33 gene (ChrX:34,437,887–34,439,090); blue: 516 bp of a long-terminal repeat with 100% identity at 9 locations in the mouse genome (e.g., ChrX:122,343,361–122,343,876).
