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Cloning of a cDNA encoding sheep calcitonin from a thyroid C-cell library
A Cumaraswamy1, M Borges, H Tamir
1Oncology Center, Johns Hopkins University School of Medicine, Baltimore, MD 21231.
Abstract:
A sheep thyroid C-cell cDNA library constructed in lambda ZAPII was screened with a probe specific for the human calcitonin (CT)-encoding gene (CT). Plaque lifts from a total of 6 x 10(4) recombinant plaques identified six overlapping cDNA clones, one of which was an 806-bp cDNA representing an almost full-length copy of the sheep CT mRNA. The nucleotide (nt) sequence consists of a 71-bp 5'-flanking sequence, a protein-coding region of 429 bp with a coding potential for a 143-amino-acid (aa) protein with a deduced M(r) of 15,888 and a 306-bp 3'-untranslated region. Northern blot analysis of sheep thyroid tissue revealed two major mRNAs with sizes of 0.9 and 0.65 kb. The sheep CT cDNA shows 71% similarity at the nt level to human CT and 58% identity at the predicted aa level.