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PMID: 6199787 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Cloning and sequence analysis of a cDNA encoding rat preprocholecystokinin.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 81 ·No. 3 ·1984-02-00 ·Pages 726-30

Deschenes RJ, Lorenz LJ, Haun RS, Roos BA, Collier KJ, Dixon JE

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 (Mr, 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 approximately equal to 750 nucleotides long. CCK mRNA of the rat brain and intestine appeared to be identical in size to the CCK mRNA of the carcinoma.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Cerebral Cortex/metabolism Cholecystokinin/genetics Cloning, Molecular DNA/isolation & purification Duodenum/metabolism Nucleic Acid Hybridization Poly A/genetics Protein Precursors/genetics RNA/genetics RNA, Messenger/genetics Rats Thyroid Neoplasms/metabolism
Chemicals
Protein Precursors RNA, Messenger Poly A RNA DNA Cholecystokinin
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Deschenes R J
Lorenz L J
Haun R S
Roos B A
Collier K J
Dixon J E
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.81.3.726
Published
1984-02-00
Pages
726-30
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC344908
Subset
IM
Grants
NIADDK NIH HHS · AM 18849 · United States
NIADDK NIH HHS · AM 21567 · United States
Databases
GENBANK
K01259
External Links
PubMed source DOI: 10.1073/pnas.81.3.726 PMC Full Text View journal details
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