A polymerase chain reaction screening strategy for the promoter of the canine dystrophin gene
Schatzberg, S., Olby, N., Steingold, S., Keene, B., Atkins, C., Meurs, K., Solomon, G., Goedegebuure, S.A., Wilton, S. and Sharp, N. (1999) A polymerase chain reaction screening strategy for the promoter of the canine dystrophin gene. American Journal of Veterinary Research, 60 (9). pp. 1040-1046.
To develop a polymerase chain reaction (PCR) strategy to screen the dystrophin promoter(s) in dogs with cardiac and skeletal myopathies.
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).
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.
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 AND CLINICAL RELEVANCE:
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.
|Publication Type:||Journal Article|
|Publisher:||American Veterinary Medical Association|
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