Journal of Aquaculture (한국양식학회지)

The Korean Society of Fisheries and Aquatic Science (한국양식학회)
권호 표지

Cloning of Growth Hormone Complementary DNA from Red-Spotted Grouper (Epinephelus akaara) and Its Expression in E. coli

붉바리(Epinephelus akaara)의 성장호르몬 cDNA의 Cloning과 E. coli에서의 발현

  • 강거영 (제주대학교 해양생물공학과) ;
  • 송춘복 (제주대학교 해양생물공학과) ;
  • 이제희 (제주대학교 해양생물공학과)
  • Published : 2003.05.01
Download PDF
⟨ Previous Next ⟩

Abstract

We have cloned and sequenced the cDNA encoding growth hormone (GH) from pituitary poly(A)$^{+}$ RNA of red-spotted grouper (Epinephelus akaara). The cDNA of red-spotted grouper GH is 883 base pairs (bp) consisting of 21 bp of 5'untranslated region (UTR), 615 Up of an open reading frame (ORF) and 247 Up of 3'UTR. The polyadenylation signal, AATAAA, was 20 bp upsteam of polyadenylation site. Based on the nucleotide sequences, the deduced putative polypeptide contains 204 amino acids (aa), representing 17 aa of a signal and 187 aa of a mature polypeptide. The putative GH cDNA encodes a polypeptide with four cysteine residues and only one N-gly- cosylation site. Comparative sequence alignment shows that red-spotted grouper GH exhibits high similarity with its corresponding other Perciformes species GH cDNAs.

붉바리(E. akaara)의 뇌하수체에서 추출한 mRNA로부터 RACE 방법으로 cDNA를 cloning하고 염기서열을 분석하였다. 붉바리의 성장호르몬 cDNA는 5' UTR, open reading frame, 3' UTR이 각각 21 bp, 615 bp, 247 bp인 전체 883 bp로 구성되어있다. Adenylation signal로 작용하는 sequence인 AATAAA는 poly(A)로부터 20 bp upstream에 위치하는 것을 확인하였다. 염기서열을 바탕으로 추정한 성장호르몬은 204개의 아미노산으로 구성된 단백질로서 signal peptide(17 aa)와 mature protein(187 aa)을 포함하고 있으며 mature protein의 분자량은 21.3 kDa으로 나타났다. 이 호르몬은 단백질의 3차 구조에 중요한 이황화 결합을 할 수 있는 4개의 cysteine 잔기와 1개의 N-glycosylation site를 가지고 있었다. 붉바리 성장호르몬의 염기서열은 농어목에 속하는 다른 어류의 성장호르몬 염기서열과의 유사도가 높게 나타났으며 orange-spotted grouper, gilthead seabream과 각각 96.9%, 88.6%의 상동성을 보였다.

Keywords

References

  1. Can. J. Fish. Aquat. Sci. v.45 Promotion of rapid growth of rainbow trout (Salmo gairdneri) by a recomibnant fish growth hormone Agellon, L. B.;C. J. Emery;J. M. Jones;S. L. Davies;A. D. Dingle;T. T. Chen https://doi.org/10.1139/f88-016
  2. Proc. Natl. Acad. Sci. USA v.84 Structure of a fish (rainbow trout) growth hormone gene and its evolutionary implications Agellon, L. B.;S. L. Davies;T. T. Chen;D. A. Powers
  3. Genome v.43 Genomic structure and sequence of the gilthead seabream (Sparus aurata) growth hormone-encoding gene: Identification of minisatellite polymorphism in intron I Almuly, R.;B. Cavari;H. Ferstman;O. Kolodny;B. Funkenstein https://doi.org/10.1139/gen-43-5-836
  4. J. Biosci. v.26 Cloning, sequncing and expression of cDNA encoding grwoth hormone from indian catfish (Heteropneustes fossilis) Anathy, V.;T. Venugopal;R. Koteeswaran;T. J. Pandian;S. Mathavan https://doi.org/10.1007/BF02703740
  5. Gen. Comp. Endocrionol. v.117 no.2 Isolation, cDNA cloning, and growth promoting activity of rabbitfish (Siganus guttatus) growth hormone Ayson, F. G.;E. G. de Jesus;Y. Amemiya;S. Moriyama;T. hirano;H. Kawauchi https://doi.org/10.1006/gcen.1999.7409
  6. Fish Physiol. Biochem. v.17 The biology of salmon growth hormone: from daylight to dominance Bjornsson, B. Th
  7. Biochemistry v.30 A heuristic approach to predicting the tertiary structure of bovine somatotropin Carlacci, L.;K. C. Chou;G. M. Maggiora https://doi.org/10.1021/bi00232a004
  8. Biochim. Biophys. Acta. v.1087 The complete nucleotide sequence of the growth-hormone gene from the common carp (Cyprinus carpio) Chiou, C. S.;H. T. Chen;W. C. Chang https://doi.org/10.1016/0167-4781(90)90126-M
  9. Fish Physiology 8 Structure and evolution of fish growth hormone and insulin-like growth factor genes Chen, T. T.;A. Marsh;M. Shamblott;K. M. Chan;Y. L. Tang;C. M. Cheng;B. Y. Yang;W. S. Hoar(eds.)(et al.)
  10. Fish Physiol. Biochem. v.17 The role of growth in endocrine regulation of salmon smoltification Dickhoff, W. W.;B. R. Beckman;D. A. Larsen;C. Duan;S. Moriyama https://doi.org/10.1023/A:1007710308765
  11. DNA Seq. v.3 Cloning and sequencing of European sea bass (Dicentrarchus labrax L.) growth hormone cDNA using polymerase chain reaction and degenerate oligonucleotides Doliana, R.;M. Bortolussi;L. Colombo https://doi.org/10.3109/10425179209034015
  12. Bio/Technology v.10 Growth enhancement in transgenic Atantic salmon by the use of an ''all fish" chimeric growth hormone gene construct Du, S. J.;Z. Gong;G. L. Fletcher;M. A.S hears;M. J. King;D. R. Idler;C. L. Hew https://doi.org/10.1038/nbt0292-176
  13. Gene v.103 Cloning and sequencing of the gilthead seabream (Sparus aurata) growth hormone-encoding cDNA Funkenstein, B.;T. T. Chen;D. A. Powers;B. Cavari https://doi.org/10.1016/0378-1119(91)90280-O
  14. J. Endocrinol. v.131 Growth regulation in the gobiid teleost, Gillichthys mirabilis: roles of growth hormone, hepatic growth hormone receptors and insulin-like growth factor-I Gray, E. S.;K. M. Kelley https://doi.org/10.1677/joe.0.1310057
  15. Gene v.77 The complete nucleotide sequence of the growth-hormone gene from Atlantic salmon (Salmo salar) Johansen, B.;O. C. Johnsen;S. Valla https://doi.org/10.1016/0378-1119(89)90079-6
  16. Gen. Comp. Endocrinol. v.101 Cloning and sequencing of the goldfish growth hormone cDNA Mahmoud, S. S.;M. M. Moloney;H. R. Habibi https://doi.org/10.1006/gcen.1996.0015
  17. Endocrinol. Rev. v.4 Structure and evolution of the growth hormone gene family Miller, W. L.;N. L. Eberhardt https://doi.org/10.1210/edrv-4-2-97
  18. Nucleic Acids Res. v.16 Nucleotide sequence of cDNA encoding the pregrowth hormone of red sea bream (Pagrus major) Momota, H.;R. Kosugi;H. Hiramatsu;H. Ohgai;A. Hara;H. Ishioka https://doi.org/10.1093/nar/16.7.3107
  19. Biosci. Biotech. Biochem. v.64 Growth regulation by insulin-like growth facotr-1 in fish Moriyama, S.;F. G. Ayson;H. Kawauchi https://doi.org/10.1271/bbb.64.1553
  20. Transgenic Res. v.10 Dramatically accelerated growth and extraordinary gigantism of transgenic mud loach Misgurnus mizolepis Nam, Y. K.;J. K. Noh;Y. S. Cho;K. N. Cho;C. G. Kim;D. S. Kim https://doi.org/10.1023/A:1016696104185
  21. Proc. Natl. Acad. Sci. USA v.68 Sequences of pituitary and placental lactogenic and growth hormones: evolution from a primordial peptide by gene reduplication Niall, H. D.;M. L. Hogan;R. Sauer;I. Y. Roshnblum;KF. C. Greenwood https://doi.org/10.1073/pnas.68.4.866
  22. DNA Seq. v.5 no.2 Cloning and sequencing of dophphifish (Coryphaena hippurus, Coryphaenidae) growth hormone-encoding cDNA Peduel, A. D.;A. Elizur;W. Knibb https://doi.org/10.3109/10425179409039713
  23. Biochemistry and Molecular Biology of Fishes 12 Growth hormone, prolactin and somatolactin: a structural overview Rand-Weaver, M.;H. Kawauchi;P. W. Hochachka(eds.);T. P. Mommsen
  24. Gen. Comp. Endocrinol. v.106 Osmoregulatory Actions of Growth Hormone and Prolactin in an Advanced Teleost Sakamoto, T.;B. S. Shepherd;S. S. Madsen;R. S. Nishioka;K. Siharath;N. H. Richman III;H. A. Bern;E. G. Grau https://doi.org/10.1006/gcen.1996.6854
  25. Biochem. Biophys. Acta. v.949 Molecular cloning and nucleotide sequence of tuna growth hormone cDNA Sato, N.;K. Watanabe;K. Murata;M. Sakaguchi;Y. Kariya;S. Kimura;M. Nonaka;A. Kimura https://doi.org/10.1016/0167-4781(88)90051-6
  26. Anal. Biochem. v.227 Revolutions in rapid amplification of cDNA ends: New strategies for polymerase chain reaction cloning of full-lengh cDNA ends Schaefer, B. C. https://doi.org/10.1006/abio.1995.1279
  27. Mol. Mar. Biol. Biotechnol. v.1 Molecular cloning and sequence analysis of the cDNA for northern pike (Esox lucius) growth hormone Schneider, J. F.;S. H. Myster;P. B. Hackett;A. Faras
  28. DNA v.1 The human growth hormone gene family: nucletide sequences show recent divergence and predict a new polypeptide hormone Seeburg, P. H. https://doi.org/10.1089/dna.1.1982.1.239
  29. DNA v.2 Efficient baterial expression of bovine and porcine growth hormones Seeburg, P. H.;S. Sias;J. P. Adelman;H. A. de Boer;J. Hayflick;P. Jhurani;D. V. Goeddel;H. L. Heyneker https://doi.org/10.1089/dna.1.1983.2.37
  30. Nature v.372 The X-ray structure of a growth hormone-prolactin receptor complex Somers, W.;M. Ultsch;A. M. D. Vos;A. A. Kossiakoff https://doi.org/10.1038/372478a0
  31. Gene v.165 Sequence of the flounder (Paralichtys olivaceus) growth hormone-encoding gene and its promoter region Tanaka, M.;Y. Toma;T. Ohkubo;S. Subo;K. Nakashima https://doi.org/10.1016/0378-1119(95)00532-B
  32. Nucleic Acids Res. v.22 CLUSTALW: improving the sensitivity of progressive multiple sequence alignment through sequence weihting, positions-specific gap penalties and weight matrix choice Thompson, J. D.;D. G. Higgins;T. J. Gibson https://doi.org/10.1093/nar/22.22.4673
  33. Comp. Biochem. Physiol. B. v.104 Molecular cloning and expression of yellowfin porgy (Acanthopagrus latus houttuyn) growth hormone cDNA Tsai, H. J.;K. L. Lin;T. T. Chen https://doi.org/10.1016/0305-0491(93)90216-R
  34. Gene v.187 Genomic structure and sequence of the pufferfish (Fugu rubripes) growth hormone-encoding gene: a comparative analysis of teleost growth hormone genes Venkatesh, B.;S. Brenner https://doi.org/10.1016/S0378-1119(96)00750-0
  35. Gen. Comp. Endocrinol. v.70 cDNA cloning and primary structure of yellow tail (Seriola quinqueradiata) pregrowthhormone Watahiki, M.;M. Tanaka;N. Masuda;M. Yamakawa;Y. Yoneda;K. Nakashima https://doi.org/10.1016/0016-6480(88)90114-1
  36. Gene v.162 Cloning of the barramundi growth hormone-encoding gene: a comparative analysis of higher and lower vertebrate GH genes Yowe, D. L.;R. J. Epping https://doi.org/10.1016/0378-1119(95)92858-5
  37. Angew Ichthyol. v.1 Novel gene transfer into the goldfish (Grassius auratus L. 1758) Zhu, Z.;G. Li;L. He;S. Z. Chen https://doi.org/10.1111/j.1439-0426.1985.tb00408.x