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Transgenic Zebrafish Biosensor for the Detection of Cadmium and Zinc Toxicity


Affiliations
1 Fish Genetics and Biotechnology Division, ICAR – Central Institute of Fisheries Education, Mumbai 400 061, India
2 CSIR – Institute of Genomics and Integrative Biology, Mathura Road, New Delhi 110 025, India
 

A transgenic zebrafish (Danio rerio) biosensor for zinc and cadmium was developed. For this, zebrafish metallothionein promoter cloned upstream to the DsRed2 gene in Tol2 transposon vector was microinjected into one/two-celled zebrafish embryos. The F1 embryos (48 h post-fertilized) of confirmed transgenics were exposed to sub-lethal doses of Cd2+, Cu2+, Hg2+ and Zn2+ for 8 h. Reporter expression was visualized as fluorescence signal and quantified using realtime PCR system. The reporter expression increased with increasing metal ion concentration. Cadmium is the most potent inducer with 4.6-fold induction followed by zinc (2.3-fold). These zebrafish biosensors could be used as a preliminary testing tool to detect heavy metals in water bodies.

Keywords

Danio rerio, DsRed2, Fluorescence, Metallothionein, Promoter.
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  • Transgenic Zebrafish Biosensor for the Detection of Cadmium and Zinc Toxicity

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Authors

Nilambari Pawar
Fish Genetics and Biotechnology Division, ICAR – Central Institute of Fisheries Education, Mumbai 400 061, India
P. Gireesh-Babu
Fish Genetics and Biotechnology Division, ICAR – Central Institute of Fisheries Education, Mumbai 400 061, India
Sridhar Sivasubbu
CSIR – Institute of Genomics and Integrative Biology, Mathura Road, New Delhi 110 025, India
Aparna Chaudhari
Fish Genetics and Biotechnology Division, ICAR – Central Institute of Fisheries Education, Mumbai 400 061, India

Abstract


A transgenic zebrafish (Danio rerio) biosensor for zinc and cadmium was developed. For this, zebrafish metallothionein promoter cloned upstream to the DsRed2 gene in Tol2 transposon vector was microinjected into one/two-celled zebrafish embryos. The F1 embryos (48 h post-fertilized) of confirmed transgenics were exposed to sub-lethal doses of Cd2+, Cu2+, Hg2+ and Zn2+ for 8 h. Reporter expression was visualized as fluorescence signal and quantified using realtime PCR system. The reporter expression increased with increasing metal ion concentration. Cadmium is the most potent inducer with 4.6-fold induction followed by zinc (2.3-fold). These zebrafish biosensors could be used as a preliminary testing tool to detect heavy metals in water bodies.

Keywords


Danio rerio, DsRed2, Fluorescence, Metallothionein, Promoter.

References





DOI: https://doi.org/10.18520/cs%2Fv111%2Fi10%2F1697-1701