Identification of the mutation in the alkaptonuria mouse model.

Alkaptonuria (aku), an inborn error of metabolism caused by the loss of homogentisate 1, 2-dioxygenase (HGD), has been described in a mouse model created by ethylnitrosourea mutagenesis but the mutation in these mice has not previously been identified.
We used RT-PCR to amplify the Hgd cDNA from Hgd (aku)/Hgd (aku) mice.
Two products shorter than the wild-type product were amplified.
Restriction mapping and DNA sequencing were then used to identify the Hgd (aku) mouse mutation, found to be a single base change in a splice donor consensus sequence, causing exon skipping and frame-shifted products.
This base change allowed us to create a non-radioactive genotyping assay for this allele.

