Identification of the first Transgenic Aquatic Animal in Iran by PCR-Based Method and Protein Analysis

Document Type : Research Article

Authors

Agricultural Biotechnology Research Institute of Iran, Gilan Branch, Agricultural Research, Education and Extension Organization (AREEO), Rasht, Iran

Abstract

In the recent years, there is evidence of training a red type of zebrafish which differs from wild-type in body color. There is not any document how it reaches to the ornamental fish farms of Iran but at first, it was a doubt it belongs to a morphotype or genetic modification (GM). First of all, a set primer was designed to validate zebrafish species. Mitochondrial 16srDNA was selected and amplified. The validation of 16S rDNA gene sequences data detected three haplotypes deposited in the GenBank. By seeking scientific articles about transgenic zebrafish, the first attention went to Glowfish (color transgenic zebrafish). We decided to build a foundation of PCR based detection for finding a gene correspondence red color of zebrafish. This sequence is specific for producing a red color in the sea anemone and it was not in genome database of wild zebrafish. A set primer was designed according to the red color of sea anemone (dsRed) to make a fragment of 680 bp in red zebrafish and cream zebrafish specimens (they had red parents). Fragment sequencing (ds/Red) revealed that it belongs to the sea anemone and none of the samples nucleotides differ from each order and positive sequence from a plasmid containing dsRed. Also, protein samples from cream and red zebrafish were extracted and evaluated with fluorescence microscope by TxRed control color. A dark and a red spread was detected for protein smear of the cream and redfish respectively. A dark spread was detected for cream fish protein smear but a red smear was marked for redfish protein. This research simply demonstrates the existence of transgenic zebrafish in Iranian ornamental fish industry.

Keywords

References

Gong Z, Wan H, Tay TL, Wang H, Chen M, Yan T. 2003. Development of transgenic fish for ornamental and bioreactor by strong expression of fluorescent proteins in the skeletal muscle. Biochem Biophys Res Commun 308: 58-63.
Waltz E. 2016. GM salmon declared fit for dinner plates. Nat Biotechnol 34: 7-9.
Du SJ, Gong Z, Fletcher GL, Shears MA, King MJ, Idler DR, Hew CL. 1992. Growth enhancement in transgenic Atlantic salmon by the use of an “all fish” chimeric growth hormone gene construct. Biotechnology 10: 176-181.
Butler T, Fletcher G. 2009. Promoter analysis of a growth hormone transgene in Atlantic salmon. Theriogenology 72: 62-71.
Chiang CY, Chen YL, Tsai HJ. 2014. Different visible colors and green fluorescence were obtained from the mutated purple chromoprotein isolated from sea anemone. Mar Biotechnol 16: 436-446.
Zeng Z, Liu X, Seebah S, Gong Z. 2005. Faithful expression of living color reporter genes in transgenic medaka under two tissue‐specific zebrafish promoters. Dev Dyn 234: 387-392.
Gong Z, He J, Ju B, Lam TJ, Xu Y, Yan T. 2006. Chimeric gene constructs for generation of fluorescent transgenic ornamental fish. The national University of Singapore.
Wan H, He J, Ju B, Yan T, Lam TJ, Gong Z. 2002. Generation of two-color transgenic zebrafish using the green and red fluorescent protein reporter genes gfp and rfp. Mar Biotechnol 4:146-154.
Rehbein H, Bogerd J. 2007. Identification of Genetically Modified Zebrafish (Danio rerio) by Protein- and DNA-Analysis. J Verbr  Lebensm 2: 122-125.
Ofelio C, Cariani A, Trentini M, Guarniero I. 2012. Novel PCR-based assay for rapid identification of Red Fluorescent Proteins in GloFish and GloFish x wildtype zebrafish (Danio rerio) hybrids. Ital J Zool 79: 541-546.
Plan D, Van den Eede G. 2010. The EU legislation on GMOs. An overview. Rome: Europea Union.
 
Journal of Genetic Resources
Volume 2, Issue 2
September 2016
Pages 93-97

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