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. 1985 Jul;82(13):4306–4310. doi: 10.1073/pnas.82.13.4306

Cloning and expression of cDNA for salmon growth hormone in Escherichia coli

Susumu Sekine *, Tamio Mizukami *, Tatsunari Nishi *, Yoshihisa Kuwana *, Akiko Saito *, Moriyuki Sato *, Seiga Itoh *,†, Hiroshi Kawauchi ‡
PMCID: PMC390402  PMID: 16593578

Abstract

cDNA clones encoding chum salmon (Oncorhynchus keta) growth hormone (sGH) have been isolated from a cDNA library prepared from chum salmon pituitary gland poly(A)+ RNA. Synthetic oligodeoxynucleotide mixtures based on amino acid residues 23-28 of sGH were used as hybridization probes to select recombinant plasmids carrying the sGH coding sequence. The complete nucleotide sequence of sGH cDNA has been determined. The cDNA sequence codes for a polypeptide of 210 amino acids, including a putative signal sequence of 22 amino acids. The 5′ and 3′ untranslated regions of the message were 64 and 426 bases long, respectively. Mature sGH was efficiently expressed in Escherichia coli carrying a plasmid in which the sGH cDNA was under control of the E. coli trp promoter; sGH comprised about 15% of the total cellular protein in such bacteria. The partially purified sGH from E. coli stimulated the growth of rainbow trout and the activity was indistinguishable from that of natural sGH.

Keywords: nucleotide, amino acid sequences, bacterial expression, growth-promoting activity

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Selected References

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  1. Cook A. F., Wilson S. W., Peter R. E. Development and validation of a carp growth hormone radioimmunoassay. Gen Comp Endocrinol. 1983 Jun;50(3):335–347. doi: 10.1016/0016-6480(83)90253-8. [DOI] [PubMed] [Google Scholar]
  2. Cooke N. E., Coit D., Weiner R. I., Baxter J. D., Martial J. A. Structure of cloned DNA complementary to rat prolactin messenger RNA. J Biol Chem. 1980 Jul 10;255(13):6502–6510. [PubMed] [Google Scholar]
  3. Farmer S. W., Hayashida T., Papkoff H., Polenov A. L. Characteristics of growth hormone isolated from sturgeon (Acipenser güldenstädti) pituitaries. Endocrinology. 1981 Feb;108(2):377–381. doi: 10.1210/endo-108-2-377. [DOI] [PubMed] [Google Scholar]
  4. Farmer S. W., Papkoff H., Hayashida T., Bewley T. A., Bern H. A., Li C. H. Purification and properties of teleost growth hormone. Gen Comp Endocrinol. 1976 Sep;30(1):91–100. doi: 10.1016/0016-6480(76)90070-8. [DOI] [PubMed] [Google Scholar]
  5. Goeddel D. V., Heyneker H. L., Hozumi T., Arentzen R., Itakura K., Yansura D. G., Ross M. J., Miozzari G., Crea R., Seeburg P. H. Direct expression in Escherichia coli of a DNA sequence coding for human growth hormone. Nature. 1979 Oct 18;281(5732):544–548. doi: 10.1038/281544a0. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Kawauchi H., Abe K., Takahashi A., Hirano T., Hasegawa S., Naito N., Nakai Y. Isolation and properties of chum salmon prolactin. Gen Comp Endocrinol. 1983 Mar;49(3):446–458. doi: 10.1016/0016-6480(83)90208-3. [DOI] [PubMed] [Google Scholar]
  8. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  11. Messing J., Crea R., Seeburg P. H. A system for shotgun DNA sequencing. Nucleic Acids Res. 1981 Jan 24;9(2):309–321. doi: 10.1093/nar/9.2.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Messing J., Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. doi: 10.1016/0378-1119(82)90016-6. [DOI] [PubMed] [Google Scholar]
  13. Miller W. L., Martial J. A., Baxter J. D. Molecular cloning of DNA complementary to bovine growth hormone mRNA. J Biol Chem. 1980 Aug 25;255(16):7521–7524. [PubMed] [Google Scholar]
  14. Niall H. D., Hogan M. L., Sauer R., Rosenblum I. Y., Greenwood F. C. Sequences of pituitary and placental lactogenic and growth hormones: evolution from a primordial peptide by gene reduplication. Proc Natl Acad Sci U S A. 1971 Apr;68(4):866–870. doi: 10.1073/pnas.68.4.866. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Niall H. D., Hogan M. L., Tregear G. W., Segre G. V., Hwang P., Friesen H. The chemistry of growth hormone and the lactogenic hormones. Recent Prog Horm Res. 1973;29:387–416. doi: 10.1016/b978-0-12-571129-6.50014-2. [DOI] [PubMed] [Google Scholar]
  16. Okayama H., Berg P. A cDNA cloning vector that permits expression of cDNA inserts in mammalian cells. Mol Cell Biol. 1983 Feb;3(2):280–289. doi: 10.1128/mcb.3.2.280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Seeburg P. H., Shine J., Martial J. A., Baxter J. D., Goodman H. M. Nucleotide sequence and amplification in bacteria of structural gene for rat growth hormone. Nature. 1977 Dec 8;270(5637):486–494. doi: 10.1038/270486a0. [DOI] [PubMed] [Google Scholar]
  20. Seeburg P. H., Sias S., Adelman J., de Boer H. A., Hayflick J., Jhurani P., Goeddel D. V., Heyneker H. L. Efficient bacterial expression of bovine and porcine growth hormones. DNA. 1983;2(1):37–45. doi: 10.1089/dna.1.1983.2.37. [DOI] [PubMed] [Google Scholar]
  21. Shine J., Seeburg P. H., Martial J. A., Baxter J. D., Goodman H. M. Construction and analysis of recombinant DNA for human chorionic somatomammotropin. Nature. 1977 Dec 8;270(5637):494–499. doi: 10.1038/270494a0. [DOI] [PubMed] [Google Scholar]
  22. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  23. Wallace R. B., Johnson M. J., Hirose T., Miyake T., Kawashima E. H., Itakura K. The use of synthetic oligonucleotides as hybridization probes. II. Hybridization of oligonucleotides of mixed sequence to rabbit beta-globin DNA. Nucleic Acids Res. 1981 Feb 25;9(4):879–894. doi: 10.1093/nar/9.4.879. [DOI] [PMC free article] [PubMed] [Google Scholar]

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