Fig. 2.

The fgt mutation of Ap1g1 and its consequences. a DNA sequencing chromatograms of control B6 +/+ and mutant fgt/fgt mice showing that the fgt mutation is a 6 bp deletion in the Ap1g1 gene. b The six base pairs deleted in the fgt mutation (shown in red) are in exon 13 of the NM_009677 reference sequence. The exon sequence is shown in capital letters and flanking intron sequences are shown in small case, italic font. Amino acids are shown below the second nucleotide of the corresponding codons. The in-frame 6 bp deletion is predicted to eliminate the phenylalanine (F) and leucine (L) amino acids shown in red. Underlined sequences in blue font correspond to the PCR primers used for mutation sequence confirmation and genotyping, and underlined sequences in green font correspond to the PCR primers used to examine the conformational effect of the two 7 bp inverted repeat sequences (double underlined). c PCR genotyping results using the blue underlined primer sequences shown in b. Ap1g1 genotypes for +/+ B6 control (lane 1), fgt/fgt (lanes 2 and 4) and +/fgt (lane 5) mice are shown with reference to 100 and 200-bp size markers (lane 3). An extra heteromeric band can be seen in the +/fgt genotype of lane 5. d PCR results using the green underlined primers shown in B, which flank both inverted repeat sequences were predicted to amplify a 207 bp product in +/+ DNA (lane 2) and a 201 bp product in fgt/fgt DNA (lane 3), with reference to 200 and 300 bp size markers (lane 1). The large extra band seen in lane 2 but absent from lane 3 is a conformational isoform of the wild-type PCR product, presumably a consequence of the two inverted repeat sequences