Untersuchungen zu familiären und rassespezifischen ...

Materials and Methods
Animals
Blood samples were collected from 17 Norwegian lundehunds representing about
one third of the German population. Four of these animals were affected by the
typical GE causing loss of proteins. Seven dogs were male, ten were female. The
group of the affected dogs consisted of three females and one male. Fourteen
lundehunds were bred in Germany, two dogs in Switzerland and one dog in The
Netherlands. All 17 dogs were related to each other as the pedigree shows (Figure
1).
All 17 lundehunds were used for analysis of the 33 bp deletion and the c.3G>A
mutation in the AMN gene as well as for genotyping microsatellite markers. Exonic
sequences and their flanking regions of the AMN gene were sequenced in four
affected and four non-affected dogs. In addition, we screened these eight dogs for
five AMN flanking SNPs described in the Broad Institute SNP collection.
Mutation Analysis
Testing for the c.1113-1145del in AMN (NC_006590.1) previously described in Giant
Schnauzers was performed by PCR-amplification using the primers 5’-
TCCGTTGCAGGCGAAGCCCCTC-3’ and 5’-CTCGGGGTGCGTGGAACCTAG-3’
and the ThermalAce DNA Polymerase kit (Invitrogen, Karlsruhe, Germany) according
to the manufacturer’s instructions.
Separation of PCR-products was performed by electrophoresis in gels containing 2%
agarose.