| Summary: | 碩士 === 國立臺灣海洋大學 === 環境生物與漁業科學學系 === 93 === The metal elements in environment, especially non-essential metals, for example cadmium, can store and cause serious damage to aquatic organisms and human by food chain. This kind of non-essential metals can induce metallothionein during the detoxification process in the organisms. Therefore, the metallothionein can be a biomarker for metal pollution. In this thesis, non-essential metal of cadmium was used to induce metallothionein and then the metallothionein induced in tilapia was separated by NuPAGE gel electrophoresis. Moreover, the fluorogenic derivatization of metallothionein with SBD-F and the specifically reacted of antibody to antigen of western blotting assay were used for analyzing the metallothionein induced in tilapia. As the results, there were a lot of sulfhydryl-rich proteins in the tissue extracts of tilapia that the crude extracts could not be analyzed accurately by SBD-F derivatization method. As results of the analysis by the production of anti-tilapia metallothionein peptide antibodies, the antibody of C-terminal peptide of tilapia metallothionein has specifically reacted with metallothionein in liver, but not in gill of tilapia. Also, none of the cadmium-induced metallothionein has been observed in the gill. Furthermore, the metallothionein is not induced in liver of tilapia under 0.5 mg/L of cadmium. In conclusion, the specifically reacted of produced antibody to metallothionein can provide a faster and convenient method on metallothionein assay.
Key words: cadmium、metallothionein、antibody、tilapia、NuPAGE gel electrophoresis、SBD-F、Western blotting。
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