Mohan-Ram, Vidyanand Sagar; (1996) The study of canine myosin heavy chain isoforms and the classification of canine breeds according to DNA fingerprinting profiles. Doctoral thesis (Ph.D), UCL (University College London).

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Abstract

The canine species presents a unique source of anatomical and athletic diversity. The evolution of the domestic dog is well documented, as are the breeding programmes which have given rise to breed-specific traits. The aims of this thesis were to apply emerging molecular biology techniques to investigate if breed-specific markers or genes could be identified between unrelated dogs. Two approaches were taken, the generation of random DNA markers by the polymerase chain reaction (PCR) and the study of specific genes, hi this last respect, canine myosin heavy chain genes (MyHC) were characterized as these encode the proteins responsible for determining the contractile properties of a muscle. The analysis of the nucleotide and primary amino acid sequences of the isolated clones yielded isoform-specific data within the locus encoding the enzymatic site of the myosin subfragment-1 domain. These have been useful in the design of specific probes. The sequence information was also used to model the canine nucleotide binding pocket and hydrolysis site to the crystal structure of the chicken pectoral MyHC isoform. The analysis of the hypervariable loop between canine isoforms revealed significant properties which could influence the rate at which adenosine triphiosphate (ATP) is utilized. During the analysis of Greyhound muscle, a novel myosin heavy chain gene was isolated and has been characterized. The random genetic analyses involved generation of DNA profiles of different breeds of dog which could confer breed-specificity when compared. In addition, part of the canine mitochondrial genome was analyzed and nucleotide differences identified between three breeds of dog. The combination of both random and specific analyses do not only have the potential to distinguish between breeds, but can also be applied to the generation of disease-markers and genome mapping efforts.

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