Cloning and sequence analysis of a cDNA encoding rat preprocholecystokinin

R. J. Deschenes, L. J. Lorenz, R. S. Haun, B. A. Roos, K. J. Collier, J. E. Dixon

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Abstract

Poly(A) RNA was isolated from a rat medullary thyroid carcinoma that exhibited high levels of immunoreactive cholecystokinin (CCK). Double-stranded cDNA was synthesized from the poly(A) RNA and inserted into the Pst I site of pBR322. Bacterial colonies containing CCK cDNA were identified using the hybridization probe d(T-C-C-A-T-C-C-A-N-C-C-C-A-T-G-T-A-G-T-C). The sequence of the probe was deduced from the known amino acid sequence of porcine CCK-8, Asp-Tyr-Met-Gly-Trp-Met-Asp-Phe-NH2. The nucleotide sequence of the cDNA complementary to the mRNA of rat preprocholecystokinin was determined. The cDNA contains 33 nucleotides in the 5'-noncoding region, 199 nucleotides in the 3'-noncoding region, and 345 nucleotides coding for a precursor to CCK, which is 115 amino acids (M(r), 12,826). Examination of the rat CCK gene revealed a suggested transcriptional control sequence analogous to the 'TATA' sequence located 33 nucleotides upstream from a proposed transcriptional start site. The amino acid sequence of CCK-39 is flanked by both amino-terminal and carboxyl-terminal extensions. Analysis of CCK mRNA showed that it is ≃750 nucleotides long. CCK mRNA of the rat brain and intestine appeared to be identical in size to the CCK mRNA of the carcinoma.

Original languageEnglish (US)
Pages (from-to)726-730
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume81
Issue number3 I
DOIs
StatePublished - 1984

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