Molecular analysis and chromosomal assignment of the canine CALC-I/α-CGRP gene
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We have isolated a recombinant phage harboring the canine CALC-I/α-CGRP gene. The gene spans a region of approx. 5.3 kb and consists of six exons with sizes ranging from 95 bp (exon 2) and 494 bp (exon 4). By alternative splicing, two transcripts with ORFs of 390 and 384 nt are generated. These encode either the 32-amino acid-long hormone calcitonin (CALC) or the neurotransmitter calcitonin gene-related peptide (α-CGRP) with a length of 37 amino acids after proteolytic processing of precursor molecules. The canine calcitonin precursor consists of 130 amino acids with a molecular mass of 14.05 kDa and a statistical pI of 8.0, whereas the deduced α-CGRP precursor harbors 128 amino acids with a molecular mass of 13.87 kDa and a statistical pI of 8.6. Both polypeptides have a common N-terminal region of 76 amino acids that is encoded by exons 2 and 3 and separated by different eight (CALC) or six (α-CGRP) amino acid spacers from the biologically active polypeptide. The CALC-I/α-CGRP gene is a member of the calcitonin gene family and was assigned to chromosome CFA 16q25.1. A comparative analysis of different dog breeds revealed a breed-specific allelic d(CAGGAG)-hexanucleotide expansion in exon 3. This expansion results in an elongation of the common N-terminal region by two amino acids (glutamine-glutamic acid) and alters the molecular mass to 14.31 kDa (pI 7.9) and 14.13 kDa (pI 8.5) of the calcitonin and α-CGRP precursor, respectively.
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