Cloning and Expression of the Cyclooxygenase-2 gene in the Rock bream, Oplegnathusfasciatus

돌돔, Oplegnathus fasciatus의 Cyclooxygenase-2 유전자의 cloning 및 발현분석

  • Jin, Ji Woong (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Kim, Do Hyung (Department of Aqualife Medicine, Chonnam National University) ;
  • Kim, Young Chul (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Jeong, Hyun Do (Department of Aquatic Life Medicine, Pukyong National University)
  • 진지웅 (부경대학교 수산생명의학과) ;
  • 김도형 (전남대학교 수산생명의학과) ;
  • 김영철 (부경대학교 수산생명의학과) ;
  • 정현도 (부경대학교 수산생명의학과)
  • Received : 2012.11.09
  • Accepted : 2013.04.02
  • Published : 2013.04.30


Megalocytivirus is a major fish pathogen in marine aquaculture of Asian countries including Korea. Despite of many species affected by this pathogen, little is known interaction between megalocytivirus and the fish immune system. One of the cyclooxygenase isoforms, named COX-2, is playing an important role in immune regulation, and distinct from COX-1 isoform of constitutive activity. COX-2 enzyme is induced by various inflammatory signals, including injection of lipopolysaccharide or infection by pathogenic agents. We cloned COX-2 gene in rock bream using degenerated primers designed from reported sequences of other fish species in PCR followed with 5'- and 3'-end RACE-PCR. The full length of cDNA of rbCOX2 (rock bream COX-2) gene are 2655 bp and that translates into 609 amino acids. The rbCOX-2 genomic organization are found to span 10 exons separated by 9 introns. We also studied if the experimental infection of rock bream with megalocytivirus could affect the expression of COX-2 gene. When injected with LPS, expression of the COX-2 gene was reached peak level at 1 day post injection and showed 13.10 fold increased level compared with that of control. While, when injected with megalocytivirus, we were not able to find significantly increased COX-2 gene expression different from that of control. Cloned and analyzed COX-2 gene in rock bream will help to understand defence mechanisms in fish after viral infection and will also support the development of the measures for treatment and prevention of viral infection.


Supported by : 농림수산식품부


  1. Chandrasekharan, N. V., Dai, H., Roos, K. L., Evanson, N. K., Tomsik, J., Elton, T. S. and Simmons, D. L.: COX-3, a cyclooxygenase-1 variant inhibited by acetaminophen and other analgesic/antipyretic drugs: cloning, structure, and expression. Proc Natl Acad Sci USA, 99:13926-13931, 2002.
  2. Cheng, S. L., Chan, L. Y., Leung, K., To, F., Go, Y. Y., Chan, Y. H., Liew, T. and Sung, J. Y.: Expression of HBx and COX-2 in chronic hepatitis B, cirrhosis and hepatocellular carcinoma: implication of HBx in upregulation of COX-2. Modern Pathology, 17:1169-1179, 2004.
  3. Choe, K. P., Havird, J., Rose, R., Hyndman, K., Piermarini, P. and Evans, D. H.: COX2 in a euryhaline teleost, Fundulus heteroclitus: primary sequence, distribution, localization, and potential function in gills during salinity acclimation. J. Exp. Biol., 209:1696-1708, 2006.
  4. Graham, S., Jeffries, A. H. and Secombes, C.: A novel assay to detect macrophages bactericidal activity in fish. Factors influencing the killing of Aeromonas salmonicida. J. Fish. Dis., 11:389-396, 1988.
  5. Havird, J. C., Miyamoto, M. M., Choe, K. P. and Evans, D. H.: Gene duplications and losses within the cyclooxygenase family of teleosts and other chordates. Mol. Biol. Evol., 25:2349-2359, 2008.
  6. Herschman, H. R.: COX-2 blockade in cancer prevention and therapy. Humana Press, 13-32, 2003.
  7. Ishikawa, T. O. and Herschman, H. R.: Two inducible, functional cyclooxygenase-2 genes are present in the rainbow trout genome. J. Cell Biochem., 102:1486-1492, 2007.
  8. Ishikawa, T. O., Griffin, K. J. P., Banerjee, U., Herschman, H. R.: The zebrafish genome contains two inducible functional cyclooxygenase-2 genes. Biochem. Biophys. Res. Commun., 352:181-187, 2007.
  9. Jeong, J. B., Kim, H. Y., Kim, K. H., Chung, J. K., Komisar, J. L. and Jeong, H. D.: Molecular comparison of iridoviruses isolated from marine fish cultured in Korea and imported from China. Aquaculture, 255: 105-116, 2006.
  10. Joo, M., Hahn, Y. S., Kwon, M., Sadikot, R. T., Blackwell, T. S. and Christman, J. W.: Hepatitis C Virus Core Protein Suppresses NF-${\kappa}B$ Activation and Cyclooxygenase-2 Expression by Direct Interaction with $I{\kappa}B$ Kinase. J. Virol., 79:7648-7657, 2005.
  11. Kim, M. S., Hwang, Y. J., Yoon, K. J., Zenke, K., Nam, Y. K., Kim, S. K. and Kim, K. H.: Molecular cloning of rock bream (Oplegnathus fasciatus) tumor necrosis factor-${\alpha}$ and its effect on the respiratory burst activity of phagocytes. Fish Shellfish Immunol., 27:618-624. 2009.
  12. Livak, K. J. and Schmittgen, T. D.: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-delta delta C(T)) method. Methods, 25:402-408, 2001.
  13. Lu, L., Wei, L., Peng, G., Mu, Y., Wu, K., Kang, L., Yan, X., Zhu, Y. and Wu, J.: NS3 protein of hepatitis C virus regulates cyclooxygenase-2 expression through multiple signaling pathways. Virology, 371:61-70, 2008.
  14. Ray, C. A., Black, R. A., Kronheim, S. R., Greenstreet, T. A., Sleath, P. R., Salvesen, G. S. and Pickup, D. J.: Viral inhibition of inflammation: cowpox virus encodes an inhibitor of the interleukin-1 beta converting enzyme. Cell, 69:597-604, 1992.
  15. Roberts, S. B., Langenau, D. M. and Goetz, F. W.: Cloning and characterization of prostaglandin endoperoxide synthase-1 and -2 from the brook trout ovary. Mol. Cell Endocrinol., 160:89-97. 2000.
  16. Savard, M., Belanger, C., Tremblay, M. J., Dumais, N., Flamand, L., Borgeat, P. and Gosselin, J.: Infection of Primary Human Monocytes by Epstein-Barr Virus. J. Immunol., 164:6467-6473. 2000.
  17. Siwicki, A., Pozet, F., Morand, M., Terech-Majewska, E. and Bernard, D.: Pathogenesis of iridovirus: in vitro influence on macrophage activity and cytokine-like protein production in fish. Acta. Vet. Brno., 70:451-456. 2001.
  18. Zenke, K. and Kim, K. H.: Molecular cloning and expression analysis of three Mx isoforms of rock bream, Oplegnathus fasciatus. Fish Shellfish Immunol., 26:599-605, 2009.