Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1982 Apr;79(8):2475–2479. doi: 10.1073/pnas.79.8.2475

Production in Escherichia coli of biologically active secretin, a gastrointestinal hormone.

M Suzuki, S Sumi, A Hasegawa, T Nishizawa, K Miyoshi, S Wakisaka, T Miyake, F Misoka
PMCID: PMC346221  PMID: 6283529

Abstract

A synthetic gene for porcine secretin was ligated with Pst I-cleaved pBR322 by its flanking synthetic Pst I linkers to produce a fused protein consisting of an amino-terminal portion of beta-lactamase and an entire secretin molecule. The hybrid plasmid was transferred into competent Escherichia coli cells. The plasmids, which were proved in our investigation to contain the secretin gene in the desired orientation, were then screened for the production of secretin. This was shown by radioimmunoassay and gel electrophoretic analysis of the polypeptides that were synthesized in E. coli minicells transformed with the hybrid plasmids. Secretin so produced, whose carboxy terminal residue may not be amidated in contrast with natural porcine secretin, showed the same activity in stimulating pancreatic secretion in a bioassay with anesthetized rats as does natural secretin.

Full text

PDF
2475

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bitar K. N., Zfass A. M., Makhlouf G. M. Binding of secretin to plastic surfaces. Gastroenterology. 1978 Dec;75(6):1080–1082. [PubMed] [Google Scholar]
  2. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  3. Byrnes D. J., Marjason J. P. Radioimmunoassay of secretin in plasma. Horm Metab Res. 1976 Sep;8(5):361–365. doi: 10.1055/s-0028-1093614. [DOI] [PubMed] [Google Scholar]
  4. Carlquist M., Jörnvall H., Mutt V. Isolation and amino acid sequence of bovine secretin. FEBS Lett. 1981 May 5;127(1):71–74. doi: 10.1016/0014-5793(81)80343-2. [DOI] [PubMed] [Google Scholar]
  5. Chang T. M., Chey W. Y. Radioimmunoassay of secretin. A critical review and current status. Dig Dis Sci. 1980 Jul;25(7):529–552. doi: 10.1007/BF01315215. [DOI] [PubMed] [Google Scholar]
  6. Dougan G., Sherratt D. The transposon Tn1 as a probe for studying ColE1 structure and function. Mol Gen Genet. 1977 Mar 7;151(2):151–160. doi: 10.1007/BF00338689. [DOI] [PubMed] [Google Scholar]
  7. Goeddel D. V., Kleid D. G., Bolivar F., Heyneker H. L., Yansura D. G., Crea R., Hirose T., Kraszewski A., Itakura K., Riggs A. D. Expression in Escherichia coli of chemically synthesized genes for human insulin. Proc Natl Acad Sci U S A. 1979 Jan;76(1):106–110. doi: 10.1073/pnas.76.1.106. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Grossman M. I. Treatment of duodenal ulcer with secretin: a speculative proposal. Gastroenterology. 1966 Jun;50(6):912–913. [PubMed] [Google Scholar]
  9. Itakura K., Hirose T., Crea R., Riggs A. D., Heyneker H. L., Bolivar F., Boyer H. W. Expression in Escherichia coli of a chemically synthesized gene for the hormone somatostatin. Science. 1977 Dec 9;198(4321):1056–1063. doi: 10.1126/science.412251. [DOI] [PubMed] [Google Scholar]
  10. Kolts B. E., McGuigan J. E. Radioimmunoassay measurement of secretin half-life in man. Gastroenterology. 1977 Jan;72(1):55–60. [PubMed] [Google Scholar]
  11. Koshland D., Botstein D. Secretion of beta-lactamase requires the carboxy end of the protein. Cell. 1980 Jul;20(3):749–760. doi: 10.1016/0092-8674(80)90321-9. [DOI] [PubMed] [Google Scholar]
  12. Martial J. A., Hallewell R. A., Baxter J. D., Goodman H. M. Human growth hormone: complementary DNA cloning and expression in bacteria. Science. 1979 Aug 10;205(4406):602–607. doi: 10.1126/science.377496. [DOI] [PubMed] [Google Scholar]
  13. Miyoshi K., Hasegawa A., Tomiyama M., Miyake T. Chemical synthesis of 27-desamidosecretin gene by the polymer support method. Nucleic Acids Symp Ser. 1981;(10):197–200. [PubMed] [Google Scholar]
  14. Mutt V., Jorpes J. E., Magnusson S. Structure of porcine secretin. The amino acid sequence. Eur J Biochem. 1970 Sep;15(3):513–519. doi: 10.1111/j.1432-1033.1970.tb01034.x. [DOI] [PubMed] [Google Scholar]
  15. Nagata S., Taira H., Hall A., Johnsrud L., Streuli M., Ecsödi J., Boll W., Cantell K., Weissmann C. Synthesis in E. coli of a polypeptide with human leukocyte interferon activity. Nature. 1980 Mar 27;284(5754):316–320. doi: 10.1038/284316a0. [DOI] [PubMed] [Google Scholar]
  16. Nilsson A., Carlquist M., Jörnvall H., Mutt V. Isolation and characterization of chicken secretin. Eur J Biochem. 1980 Nov;112(2):383–388. doi: 10.1111/j.1432-1033.1980.tb07216.x. [DOI] [PubMed] [Google Scholar]
  17. Roozen K. J., Fenwick R. G., Jr, Curtiss R., 3rd Synthesis of ribonucleic acid and protein in plasmid-containing minicells of Escherichia coli K-12. J Bacteriol. 1971 Jul;107(1):21–33. doi: 10.1128/jb.107.1.21-33.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Shine J., Fettes I., Lan N. C., Roberts J. L., Baxter J. D. Expression of cloned beta-endorphin gene sequences by Escherichia coli. Nature. 1980 Jun 12;285(5765):456–463. doi: 10.1038/285456a0. [DOI] [PubMed] [Google Scholar]
  19. Sutcliffe J. G. Nucleotide sequence of the ampicillin resistance gene of Escherichia coli plasmid pBR322. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3737–3741. doi: 10.1073/pnas.75.8.3737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tachibana S. The bioassay of secretin in the rat. Jpn J Pharmacol. 1971 Jun;21(3):325–336. doi: 10.1254/jjp.21.325. [DOI] [PubMed] [Google Scholar]
  21. Tai H. H., Chey W. Y. Rapid extraction of secretin from plasma by XAD-2 resin and its application in the radioimmunoassay of secretin. Anal Biochem. 1978 Jul 1;87(2):376–385. doi: 10.1016/0003-2697(78)90687-5. [DOI] [PubMed] [Google Scholar]
  22. Wetzel R., Heyneker H. L., Goeddel D. V., Jhurani P., Shapiro J., Crea R., Low T. L., McClure J. E., Thurman G. B., Goldstein A. L. Production of biologically active N alpha-desacetylthymosin alpha 1 in Escherichia coli through expression of a chemically synthesized gene. Biochemistry. 1980 Dec 23;19(26):6096–6104. doi: 10.1021/bi00567a023. [DOI] [PubMed] [Google Scholar]
  23. Yanaihara N., Sakagami M., Sato H., Yammamoto K., Hashimoto T. Immunological aspects of secretin, substance P, and VIP. Gastroenterology. 1977 Apr;72(4 PT2):803–810. [PubMed] [Google Scholar]
  24. ZOLLINGER R. M., ELLISON E. H. Primary peptic ulcerations of the jejunum associated with islet cell tumors of the pancreas. Ann Surg. 1955 Oct;142(4):709-23; discussion, 724-8. [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES