Abstract
Metallothioneins (MTs) are a family of low-molecular-weight, cysteine-rich proteins that have a high affinity for divalent ions. Recently, MT level has become a major biomarker for monitoring metal pollution in fish. As a first step towards the development of a reliable and standard assay method for the detection of fish MTs in Hong Kong waters, reverse transcription-polymerase chain reaction (RT-PCR) was employed to obtain complementary DNAs (cDNAs) coding for goldfish ( Carrasius auratus) and tilapia ( Tilapia mossambica) MTs. Based on the cloned tilapia MT cDNA sequence, specific primers were designed to detect MT mRNA levels in tilapia tissues using RT-PCR. Screening of a common carp liver cDNA library also yielded MT cDNAs. For the development of an immuno-assay for monitoring MT levels in fish tissues, synthetic peptide, made according to the conserved teleost MT N-terminal amino acid sequences, was employed to raise anti-fish MT antibodies. These molecular and immunological probes will be useful in 1. testing the feasibility of using tilapia MT levels as a biomarker for monitoring metal pollution in Hong Kong fish and 2. determining the basic physiological significance of MT in metal tolerance in tilapia.
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