Isolation of the [beta]-Galactosidase Gene Responsible for GM1 Gangliosidosis in Sheep

GMl gangliosidosis is a genetic, lysosomal storage disease that occurs in cats, humans, cattle, dogs, and sheep. In humans there are three forms of the disease, infantonset, juvenile-onset, and adult-onset while in animals there is one form that is similar to the human juvenile-onset form. Gangliosidosis is caused by a defective P-galactosidase gene which results in an accumulation of the glycolipid GM1 ganglioside in cells of the body. Neuronal cells store the largest amounts of GM1 ganglioside causing serious neurological symptoms in affected individuals. Our objective was to develop a rapid test to identify gangliosidosis in-utero during the first trimester of pregnancy. We constructed primers from areas homologous between the mouse and human cDNA sequences that were also homologous in the dog amino acid sequence to use in the amplification of the sheep Pgalactosidase cDNA by PCR Human and mouse acid P-galactosidase cDNAs have been sequenced and were shown upon comparison to be ~ 70% homologous. Dog amino acid sequence was shown to be ~85% homologous to the human amino acid sequence. PCR was performed on mouse and human P-galactosidase cDNA clones, a sheep pituitary cDNA library, RNA from unaffected sheep, and DNA from unaffected sheep. We did not successfully isolate the sheep P-galactosidase cDNA due to complications with PCR reactions so a test was not developed in this research