Positional Cloning of l11Jus1 and l11Jus4. A. Exclusion Mapping Breeding Scheme. Blue line: ENU-mutated allele (l11Jus1; C57BL/6J); Green line with the double arrow: 34 Mb inversion (In(11Trp53;11Wnt3)8Brd; 129S6/SvEvTac); yellow line: dominant Rex allele (129S6/SvEvTac), which confers a curly coat. Plus sign: wild type locus. All animals carrying two copies of the l11Jus1 allele will die in utero. Representative phenotypes and crossover events are depicted in the F2 generation. B. Mapping the Nle1 mutation by haplotype analysis in the F2generation. Each box represents a locus within Mmu 11. Yellow boxes are homozygous 129S6/SvEvTac genotypes; green boxes are heterozygous for 129S6/SvEvTac and C57BL/6J genotypes; while blue boxes indicate homozygous C57BL/6J genotypes. Wsb1 &D11Mit120 define the boundaries of the critical region. C. Physical map of the non-recombinant interval. Nle1 lies ~700 kb centromeric to D11Mit120. D. Mutation analysis. Arrows indicate the location of each mutation. Stacked chromatographs show the sequence for each strain. E. The Nle1 cDNA and genotyping primers. The full-length, spliced mRNA product is shown with exons represented by empty boxes. Primer locations are depicted by arrows. The 5’ and 3’UTRs are represented by a thin line on the mRNA transcript. F. NLE1 protein structure. Green oval: NLE specific domain; yellow boxes: WD40 Repeat-like domains; green diamonds: mutation sites.
Lossie et al. BMC Genetics 2012 13:106 doi:10.1186/1471-2156-13-106