Fisheries and Aquatic Sciences

The Korean Society of Fisheries and Aquatic Science (한국수산과학회)
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CgGH insertion functional domain analysis in transgenic G1 and G2 and G3 mutiara catfish (Clarias gariepinus) broodstock

  • Buwono, Ibnu Dwi (Aquaculture Department, Faculty of Fisheries and Marine Sciences, Universitas Padjadjaran) ;
  • Grandiosa, Roffi (Aquaculture Department, Faculty of Fisheries and Marine Sciences, Universitas Padjadjaran) ;
  • Mulyani, Yuniar (Aquaculture Department, Faculty of Fisheries and Marine Sciences, Universitas Padjadjaran)
  • Received : 2021.06.22
  • Accepted : 2021.10.26
  • Published : 2022.01.30
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Abstract

Catfish is one of the most important freshwater fish farming commodities in Indonesia. Higher catfish production can be achieved by cultivating transgenic catfish carrying the growth hormone (GH) gene of African catfish (Clarias gariepinus GH, CgGH). This research focuses on analysis of the presence of the CgGH gene in transgenic G1, G2, and G3 mutiara catfish broodstock, as an indication of stable CgGH inheritance. CgGH gene was isolated using the RNeasy mini kit and RT-PCR. RT-PCR revealed amplicons measuring approximately 600 bp in transgenic G0, G1, G2, and G3 mutiara catfish. The CgGH consensus sequence similarities ranged from 93.76% to 97.06%, with four functional domain sites (somatotropin-1, somatotropin-2, four α-helix, N-glycosylation, four cysteine residues) of fish GH proteins. The functional domains of fish GH proteins are conserved in G1, G2, and G3 and indicate stable exogenous GH inheritance to produce transgenic catfish strains in each generation.

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Acknowledgement

The authors are thankful to the Directorate of Research and Community Service of the Universitas Padjadjaran for support of research. The author would like to thank to the research team and research assistants (Rahayu Ramadhayanti and Ayuniar Puteri) thank you for the technical support during the research.

References

  1. Anathy V, Venugopal T, Koteeswaran R, Pandian TJ, Mathavan S. Cloning, sequencing and expression of cDNA encoding growth hormone from Indian catfish (Heteropneustes fossilis). J Biosci. 2001;26:315-24. https://doi.org/10.1007/BF02703740
  2. Buwono ID, Iskandar, Agung MUK, Subhan U. Construction of transgenic catfish (Clarias sp.) using sperm electrophoration technique. J Biol. 2016;20:17-28.
  3. Buwono ID, Iskandar I, Grandiosa R. Growth hormone transgenesis and feed composition influence growth and protein and amino acid content in transgenic G3 mutiara catfish (Clarias gariepinus). Aquac Int. 2021;29:431-51. https://doi.org/10.1007/s10499-020-00628-8
  4. Buwono ID, Junianto J, Iskandar I, Alimuddin A. Growth and expression level of growth hormone in transgenic mutiara catfish second generation. J Biotech Res. 2019a;10:102-9.
  5. Buwono ID, Junianto J, Iskandar I, Alimuddin A. Reproduction performance of transgenic mutiara catfish (G1) comprising the growth hormone gene. J Biotech Res. 2019b;10:199-212.
  6. Degani G, Jackson K, Yom-Din S, Goldberg D. cDNA cloning and mRNA expression of growth hormone in Belontiidae (Anabantoidei suborder). Isr J Aquac Bamidgeh. 2006;58:124-36.
  7. Hew CL, Davies PL, Fletcher G. Antifreeze protein gene transfer in Atlantic salmon. Mol Mar Biol Biotechnol. 1992;1:309-17.
  8. Hinits Y, Moav B. Growth performance studies in transgenic Cyprinus carpio. Aquaculture. 1999;173:285-96. https://doi.org/10.1016/S0044-8486(98)00452-9
  9. Iwai T, Inoue S, Kotani T, Yamashita M. Production of transgenic medaka fish carrying fluorescent nuclei and chromosomes. Zool Sci. 2009;26:9-16. https://doi.org/10.2108/zsj.26.9
  10. Mori T, Devlin RH. Transgene and host growth hormone gene expression in pituitary and nonpituitary tissues of normal and growth hormone transgenic salmon. Mol Cell Endocrinol. 1999;149:129-39. https://doi.org/10.1016/S0303-7207(98)00248-2
  11. Nam YK, Cho YS, Chang YJ, Jo JY, Kim DS. Generation of transgenic homozygous line carrying the CAT gene in mud loach Misgurnus mizolepis. Fish Sci. 2000;66:58-62. https://doi.org/10.1046/j.1444-2906.2000.00008.x
  12. Nam YK, Noh JK, Cho YS, Cho HJ, Cho KN, Kim CG, et al. Dramatically accelerated growth and extraordinary gigantism of transgenic mud loach Misgurnus mizolepis. Transgenic Res. 2001;10:353-62. https://doi.org/10.1023/A:1016696104185
  13. Peyush P, Moriyama S, Takahashi A, Kawauchi H. Molecular cloning of growth hormone complementary DNA in barfin flounder (Verasper moseri). Mar Biotechnol. 2000;2:21-6. https://doi.org/10.1007/s101269900004
  14. Pinheiro JS, Wolff JLC, Araujo RC, Hilsdorf AWS. Molecular cloning and sequence analysis of growth hormone cDNA of neotropical freshwater fish pacu (Piaractus mesopotamicus). Genet Mol Biol. 2008;31:381-4. https://doi.org/10.1590/S1415-47572008000200037
  15. Rahman MA, Maclean N. Growth performance of transgenic tilapia containing an exogenous piscine growth hormone gene. Aquaculture. 1999;173:333-46. https://doi.org/10.1016/S0044-8486(98)00456-6
  16. Rajesh R, Majumdar KC. A comparative account of the structure of the growth hormone encoding gene and genetic interrelationship in six species of the genus Labeo. Fish Physiol Biochem. 2007;33:311-33. https://doi.org/10.1007/s10695-007-9164-3
  17. Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci. 1977;74:5463-7. https://doi.org/10.1073/pnas.74.12.5463
  18. Tewari R, Michard-Vanhee C, Perrot E, Chourrout D. Mendelian transmission, structure and expression of transgenes following their injection into the cytoplasm of trout eggs. Transgenic Res. 1992;1:250-60. https://doi.org/10.1007/BF02525166
  19. Uh M, Khattra J, Devlin RH. Transgene constructs in coho salmon (Oncorhynchus kisutch) are repeated in a head-to-tail fashion and can be integrated adjacent to horizontally-transmitted parasite DNA. Transgenic Res. 2006;15:711-27. https://doi.org/10.1007/s11248-006-9016-4
  20. Vaz BS, Cerqueira GM, Silva JC, Manzke VHB, Moreira CGA, Moreira HLM. Sequence analysis of the growth hormone gene of the South American catfish Rhamdia quelen. Genet Mol Res. 2010;9:2184-90. https://doi.org/10.4238/vol9-4gmr708
  21. Xiong J. 15 - Protein tertiary structure prediction. In Xiong J, editor. Essential bioinformatics. Cambridge, UK: Cambridge University Press; 2006. p. 214-30.
  22. Yang W, Zhang N, Shi B, Zhang S, Zhang L, Zhang W. Isotocin regulates growth hormone but not prolactin release from the pituitary of ricefield eels. Front Endocrinol. 2018;9:166. https://doi.org/10.3389/fendo.2018.00166
  23. Zhang M, Chen C, Guo Y, Guo J, Wang X. Cloning and sequence analysis of full-length growth hormone cDNA from Clarias gariepinus. Acta Agric Boreal Sin. 2009;24:27-32.