A polymerase chain reaction screening strategy for the promoter of the canine dystrophin gene.
OBJECTIVE
To develop a polymerase chain reaction (PCR) strategy to screen the dystrophin promoter(s) in dogs with cardiac and skeletal myopathies.


METHODS
9 Doberman Pinschers, 1 Dalmation, and 1 Saint Bernard with dilated cardiomyopathy (DCM); 1 Irish Terrier with muscular dystrophy; and 2 dystrophin-deficient German Shorthaired Pointers (GSHP).


METHODS
For each of the 3 unique exons associated with the muscle (M), Purkinje (P), and cortical (C) promoters of the dystrophin gene, each first exon, and the M promoter plus its first exon, were amplified, cloned, and sequenced. The M dystrophin transcript was amplified by reverse transcriptase PCR from skeletal and cardiac muscle RNA of 1 Doberman Pinscher and from skeletal muscle RNA of 1 GSHP.


RESULTS
The M, P, and C first exons were amplified from all dogs except the 2 GSHP, which had a deletion encompassing the entire M, P, and C dystrophin promoter region. The M transcript could not be amplified from muscles of the GSHP, but was amplified from skeletal and cardiac muscle of the Doberman Pinscher. Sequencing of the product representing the M promoter and its first exon revealed no differences between clinically normal dogs and the Doberman Pinscher with DCM.


CONCLUSIONS
We have ruled out a major rearrangement of the dystrophin promoter region as the universal cause of DCM in Doberman Pinschers or of Irish Terrier myopathy. Use of the strategy identified a large deletion of this region in muscle from the GSHP.
Schatzberg
S
S
Department of Companion Animal and Special Species Medicine, College of Veterinary Medicine, North Carolina State University, Raleigh 27606, USA.
Olby
N
N
Steingold
S
S
Keene
B
B
Atkins
C
C
Meurs
K
K
Solomon
G
G
Goedegebuure
S A
SA
Wilton
S
S
Sharp
N
N
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern / Q000662:veterinary; D002311:Cardiomyopathy, Dilated / Q000175:diagnosis / Q000235:genetics / Q000662:veterinary; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry; D017931:DNA Primers / Q000737:chemistry; D004283:Dog Diseases / Q000175:diagnosis / Q000235:genetics*; D004285:Dogs; D016189:Dystrophin / Q000737:chemistry / Q000172:deficiency / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D005091:Exons; D006801:Humans; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000737:chemistry; D009137:Muscular Dystrophy, Animal / Q000175:diagnosis / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011689:Purkinje Cells / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
D017931:DNA Primers; D016189:Dystrophin; D004247:DNA