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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
. 1983 May;80(10):2866–2869. doi: 10.1073/pnas.80.10.2866

Molecular cloning of human gastrin cDNA: evidence for evolution of gastrin by gene duplication.

E Boel, J Vuust, F Norris, K Norris, A Wind, J F Rehfeld, K A Marcker
PMCID: PMC393933  PMID: 6574456

Abstract

An oligo(dT)-primed cDNA copy of the mRNA coding for the human gastrin precursor was constructed from poly(A)-containing RNA from a human pancreatic, gastrin-producing tumor (a gastrinoma). The cDNA was inserted into the Pst I endonuclease site of plasmid pBR322 by the use of the poly(dC) and poly(dG) tailing procedure. Clones containing gastrin sequences were selected by hybridization to a purified single-stranded 32P-labeled gastrin cDNA probe. This probe was constructed with gastrinoma mRNA as template. As primer for the cDNA synthesis, we used a synthetic oligonucleotide mixture, d(AG-A-A-AG-T-C-C-A-T-C-C-A), corresponding to the gastrin-specific amino acid sequence Trp-Met-Asp-Phe. In this way we determined the nucleotide sequence of the entire coding region (303 nucleotides), the entire 3' untranslated region (102 nucleotides), and 8 nucleotides of the 5' untranslated region. A striking homology between parts of the coding region suggests that evolution of the gastrin gene has involved a gene duplication.

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Selected References

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  1. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baldwin G. S. Gastrin and the transforming protein of polyoma virus have evolved from a common ancestor. FEBS Lett. 1982 Jan 11;137(1):1–5. doi: 10.1016/0014-5793(82)80302-5. [DOI] [PubMed] [Google Scholar]
  3. Bentley P. H., Kenner G. W., Sheppard R. C. Structures of human gastrins I and II. Nature. 1966 Feb 5;209(5023):583–585. doi: 10.1038/209583b0. [DOI] [PubMed] [Google Scholar]
  4. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blobel G., Dobberstein B. Transfer of proteins across membranes. I. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma. J Cell Biol. 1975 Dec;67(3):835–851. doi: 10.1083/jcb.67.3.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Buell G. N., Wickens M. P., Payvar F., Schimke R. T. Synthesis of full length cDNAs from four partially purified oviduct mRNAs. J Biol Chem. 1978 Apr 10;253(7):2471–2482. [PubMed] [Google Scholar]
  7. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  8. Dagert M., Ehrlich S. D. Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene. 1979 May;6(1):23–28. doi: 10.1016/0378-1119(79)90082-9. [DOI] [PubMed] [Google Scholar]
  9. Dockray G. J., Vaillant C., Hopkins C. R. Biosynthetic relationships of big and little gastrins. Nature. 1978 Jun 29;273(5665):770–772. doi: 10.1038/273770a0. [DOI] [PubMed] [Google Scholar]
  10. Friedman E. Y., Rosbash M. The syntheiss of high yields of full-length reverse transcripts of globin mRNA. Nucleic Acids Res. 1977 Oct;4(10):3455–3471. doi: 10.1093/nar/4.10.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. GREGORY R. A., TRACY H. J. A NOTE ON THE NATURE OF THE GASTRIN-LIKE STIMULANT PRESENT IN ZOLLINGER-ELLISON TUMOURS. Gut. 1964 Apr;5:115–117. [PubMed] [Google Scholar]
  12. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ito H., Ike Y., Ikuta S., Itakura K. Solid phase synthesis of polynucleotides. VI. Further studies on polystyrene copolymers for the solid support. Nucleic Acids Res. 1982 Mar 11;10(5):1755–1769. doi: 10.1093/nar/10.5.1755. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Larsson L., Rehfeld J. F. A peptide resembling COOH-terminal tetrapeptide amide of gastrin from a new gastrointestinal endocrine cell type. Nature. 1979 Feb 15;277(5697):575–578. doi: 10.1038/277575a0. [DOI] [PubMed] [Google Scholar]
  15. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Noyes B. E., Mevarech M., Stein R., Agarwal K. L. Detection and partial sequence analysis of gastrin mRNA by using an oligodeoxynucleotide probe. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1770–1774. doi: 10.1073/pnas.76.4.1770. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Peacock S. L., McIver C. M., Monahan J. J. Transformation of E. coli using homopolymer-linked plasmid chimeras. Biochim Biophys Acta. 1981 Sep 28;655(2):243–250. doi: 10.1016/0005-2787(81)90014-9. [DOI] [PubMed] [Google Scholar]
  18. Proudfoot N. J., Brownlee G. G. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. doi: 10.1038/263211a0. [DOI] [PubMed] [Google Scholar]
  19. Rehefeld J. F. Three components of gastrin in human serum. Gel filtration studies on the molecular size of immunoreactive serum gastrin. Biochim Biophys Acta. 1972 Dec 28;285(2):364–372. [PubMed] [Google Scholar]
  20. Rehfeld J. F., Larsson L. I. Pituitary gastrins. Different processing in corticotrophs and melanotrophs. J Biol Chem. 1981 Oct 25;256(20):10426–10429. [PubMed] [Google Scholar]
  21. Rehfeld J. F., Larsson L. I. The predominating molecular form of gastrin and cholecystokinin in the gut is a small peptide corresponding to their COOH-terminal tetrapeptide amide. Acta Physiol Scand. 1979 Jan;105(1):117–119. doi: 10.1111/j.1748-1716.1979.tb06320.x. [DOI] [PubMed] [Google Scholar]
  22. Rehfeld J. F. Localisation of gastrins to neuro- and adenohypophysis. Nature. 1978 Feb 23;271(5647):771–773. doi: 10.1038/271771a0. [DOI] [PubMed] [Google Scholar]
  23. Rehfeld J. F., Stadil F. Gel filtration studies on immunoreactive gastrin in serum from Zollinger-Ellison patients. Gut. 1973 May;14(5):369–373. doi: 10.1136/gut.14.5.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rehfield J. F., Uvnäs-Wallensten K. Gastrins in cat and dog: evidence for a biosynthetic relationship between the large molecular forms of gastrin and heptadecapeptide gastrin. J Physiol. 1978 Oct;283:379–396. doi: 10.1113/jphysiol.1978.sp012507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Roychoudhury R., Jay E., Wu R. Terminal labeling and addition of homopolymer tracts to duplex DNA fragments by terminal deoxynucleotidyl transferase. Nucleic Acids Res. 1976 Jan;3(1):101–116. doi: 10.1093/nar/3.1.101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Soeda E., Arrand J. R., Smolar N., Walsh J. E., Griffin B. E. Coding potential and regulatory signals of the polyoma virus genome. Nature. 1980 Jan 31;283(5746):445–453. doi: 10.1038/283445a0. [DOI] [PubMed] [Google Scholar]
  27. Uvnäs-Wallensten K., Rehfeld J. F., Larsson L. I., Uvnäs B. Heptadecapeptide gastrin in the vagal nerve. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5707–5710. doi: 10.1073/pnas.74.12.5707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wood W. B. Host specificity of DNA produced by Escherichia coli: bacterial mutations affecting the restriction and modification of DNA. J Mol Biol. 1966 Mar;16(1):118–133. doi: 10.1016/s0022-2836(66)80267-x. [DOI] [PubMed] [Google Scholar]
  29. Yalow R. S., Berson S. A. Size and charge distinctions between endogenous human plasma gastrin in peripheral blood and heptadecapeptide gastrins. Gastroenterology. 1970 May;58(5):609–615. [PubMed] [Google Scholar]
  30. Yoo O. J., Powell C. T., Agarwal K. L. Molecular cloning and nucleotide sequence of full-length of cDNA coding for porcine gastrin. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1049–1053. doi: 10.1073/pnas.79.4.1049. [DOI] [PMC free article] [PubMed] [Google Scholar]

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