Development of a fluorescent transgenic zebrafish biosensor for sensing aquatic heavy metal pollution

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• 作者：Nilambari Pawar ; P. Gireesh-Babu ; Supriya Sabnis ; Kiran Rasal…
• 关键词：Tol2 transposon vector ; Fluorescent reporter ; Metallothionein ; Bioreporter ; Perna viridis
• 刊名：Transgenic Research
• 出版年：2016
• 出版时间：October 2016
• 年：2016
• 卷：25
• 期：5
• 页码：617-627
• 全文大小：937 KB
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Biomedicine
  Molecular Medicine
  Plant Genetics and Genomics
  Animal Genetics and Genomics
  Plant Sciences
  Human Genetics
  
• 出版者：Springer Netherlands
• ISSN：1573-9368
• 卷排序：25

文摘

We report a transgenic zebrafish (Danio rerio) designed to respond to heavy metals using a metal-responsive promoter linked to a fluorescent reporter gene (DsRed2). The metallothionein MT-Ia1 promoter containing metal-responsive elements was derived from the Asian green mussel, Perna viridis. The promoter is known to be induced by a broad spectrum of heavy metals. The promoter-reporter cassette cloned into the Tol2 transposon vector was microinjected into zebrafish embryos that were then reared to maturity. A transgene integration rate of 28 % was observed. The confirmed transgenics were mated with wild-type counterparts, and pools of F₁ embryos were exposed to sub-lethal doses of Cd2+, Cu2+, Hg2+, Pb2+ and Zn2+. The red fluorescence response of zebrafish embryos was observed 8 h post- exposure to these sub-lethal doses of heavy metals using a fluorescence microscope. Reporter expression estimated by real-time PCR revealed eightfold, sixfold and twofold increase on exposure to highest concentrations of Hg2+, Cd2+ and Cu2+, while Pb2+ and Zn2+ had no effect. This biosensor could be a first-level screening method for confirming aquatic heavy metal bio-toxicity to eukaryotes.