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Transfer gene technology has affected on the science progress, such in modern biology and biotechnology (Power et al. 1998). In aquaculture, transgenic technology has been applied in several fish species like tilapia (Rachman & Maclean, 1992; Martinez et al. 1996; Maclean et al. 2002), rainbow trout (Guyomard et al. 1989), Atlantic salmon (Shears et al. 1991), gold fish (Zhu et al. 1985), common carp (Hinits & Moav, 1999), zebrafish (Sheela et al. 1998; Morales et al. 2001; Jesuthasan & Subburaju, 2002), ayu (Cheng et al. 2002), silver bass (Lu et al. 2002) and medaka (Sarmasik 2002).
Although transgenesis is successful in several species, but there are some problems in finding a suitable promoter for expression of target genes. Some groups of researchers have success when using the construction that the gene source not from fish, but some other groups fail, i.e. the gene that introduced not integrated in the host genome (Chourrout et al. 1990; Penman et al. 1991). This problem can be overcome by using the promoter that has a kinship with the fish. This shows that the promoter is derived from fish are often more efficient compared with that derived from the virus in mammals and in fish transgenesis (Alam et al. 1996).
One important consideration in transgenesis technology is promoter selection; the expression of genes coding protein related to a character. Promoter constitute DNA sequence that is located on the upstream of genes (Hackett 1993; Glick & Pasternak, 2003) which will set the place, time and level of genes expression. Thus, the promoter genes as a switch and become one of the determine factors of transgenesis success.
Some of the promoter already isolated and tried on several fish species by several researchers; cytomegalovirus (CMV) from the human virus, elongation factor-1 alpha (EF-1α) of medaka, beta-actin of medaka, and myosin light chain-2 (Mylz-2) of the zebrafish. In zebrafish, b-actin promoter and Mylz-2 showed stronger activity than EF-1α and CMV. This is possibility that not all of cis-acting elements from human virus are recognized by trans-acting factors of zebrafish (Alimuddin 2003).
Based on the activity in tilapia, some promoter is very potential to be used. However, the promoters used in this research are heat shock, Japanese flounder keratin, medaka β-Actin and tilapia β-Actin. All of promoters already used on the rainbow trout, tilapia and zebra fish (Hwang et al. 2003) with high level of expression. Use of the four promoters in this research is based on the consideration that the entire promoter is derived from fish and active in tilapia (preliminary research). The four promoters are also very possible to expression not only for growth gen, but also for other important characters such as diseases resistance.
Literature
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Alimuddin. 2003. Introduction and Expression of foreign delta 6 desaturase-Like Gene in a Teleostean fish. Thesis. Graduate School of Fisheries Science. Tokyo University of Fisheries.
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Alam MS, Lavender FL, Iyengar A, Rahman MA, Ayad HH, Lathe R, Morley SD, Maclean N. 1996. Comparison of the activity of carp and rat ß-actin gene regulatory sequences in tilapia and rainbow trout embryos. Mol. Reprod. Dev, 45 : 117-122.
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Cheng CA, Lu KL, Lau EL, Yang TY, Lee CY, Wu JL and Chang CY. 2002. Growth promotion in Ayu (Plecoglossus altivelis) by gene transfer of the rainbow trout growth hormone gene. Zoological Studies 41(3):303-310.
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Chourrout DR, Guyomard R, Houdebine LM. 1990. Techniques for the development of transgenic fish: a review, in: Church RB (Ed.). Transgenic Model in Medicine and Agriculture. Wiley-Liss, New York. pp. 89-99.
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Glick BR and Pasternak JJ. 2003. Molecular biotechnology: principles and application of recombinant DNA. Third edition. ASM Press. Washington DC.
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Guyomard R, Chourrout D, Leroux C, Houdebine LM and Pourrain F. 1989. Integration and germ line transmission of foreign genes microinjected into fertilized trout eggs. Biochimie 71, 857–863.
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Hacket PB. 1993. The molecular biology of transgenic fish. In : Hocachka and Mommesen (Eds.). Biochemistry and Molecular Biology of Fishes, 2:218 - 221.
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Hinits Y and Moav B. 1999. Growth performance studies in transgenic Cyprinus carpio. Aquaculture 173: 285–296
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Hwang GL, Rahman MA, Razak SA, Sohm F, Farahmand H, Smith A, Brooks C and Maclean N. 2003. Isolation and charactrisation of tilapia ?-actin promoter and comparison of its activity with carp beta-actin promoter. Biochimica et Biophsica Acta, 1625: 11-18
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Jesuthasan S and Subburaju S. 2002. Gene transfer into zebrafish by sperm nuclear transplantation. Developmental Biology 242, 88–95
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Lu JK, Fu BH, Wu JL and Chen TT. 2002. Production of transgenic silver sea bream (Sparus sarba) by different gene transfer methods. Mar. Biotechnol. 4, 328-337
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Maclean N and Laight RJ. 2000. Transgenic fish: an evaluation of benefits and risks. Fish and Fisheries 1: 146-172
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Morales R, Herrera MT, Arenal A, Cruz A, Hernández O, Pimentel R, Guillén I, Martínez R, Estrada MP. 2001. Tilapia chromosomal growth hormone gene expression accelerates growth in transgenic zebrafish (Danio rerio). EJB Electronic Journal of Biotechnology. ISSN: 0717-3458 Vol.4 No.2, Issue of August 15. This paper is available on line at http://www.ejb.org/content/vol4/issue2/full/3
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Penman DJ, Iyanger A, Beeching AJ, Rahman A, Sulaiman Z, Maclean N. 1991. Pattern of transgene inheritance in rainbow trout (Onchorynchus mykiss). Mol. Reprod. Dev. 30: 201-206.
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Powers DA, Gómez CM, Chen TT and Dunham RA. 1998. Genetic Enginering of Finfish and shellfish. In: de la Fuente J. and Castro F.O. eds. Gene transfer in aquatic organisms. RG Landes Company and Germany, Springer-Verlag, Austin, Texas, USA. pp. 17-34.
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Rahman MA and Maclean N. 1992. Production of transgenic tilapia (Oreochromis niloticus) by one-cell- stage microinjection. Aquaculture, 105: 219-232.
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Shears, M.A., Fletcher, G.L., Hew, C.L., Gauthier, S. and Davies, P.L. (1991) Transfer, expression, and stable inheritance of antifreeze protein genes in Atlantic salmon (Salmo salar). Molecular Marine Biology and Biotechnology 1, 58–63.
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Zhu Z, Li G, He L and Chen S. 1985. Novel gene transfer into the fertilized eggs of goldfish (Carassius auratus 1758). Journal of Applied Ichthyology 1, 31–33.
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Sheela SG, Chen JD, Mathavan S and Pandian TJ. 1998. Construction, electroporatic transfer and expression of Zp?JypGH and Zp?JrtGH in zebrafish. J. Biosci., 23, No.5, pp 565-576.
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Sarmasik, A. 2003. Application of Gene Transfer Technology for Genetic Improvement of Fish. Turk J Zool 27 (2003) 1-6
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