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(P043) Vitellogenin mRNA induction in crucian carp (Carassius auratus) by estrogenic chemicals.
Park, Eung-Roh*,1, Jeon, Seong-Hwan1, Na, Jin-Gyun1, Park, Kwangsik1, 1 National Institute of Environmental Research, Gyeongseo-dong, Seo-gu, Incheon
ABSTRACT- Vitellogenin is a precursor of egg-yolk protein and secreted by the liver of female as well as male fish in response to estrogenic compounds, so the protein induction means the exposure of endocrine disrupting chemcals. Vitellogenin production was measured by ELISA using monoclonal antibody or by mRNA expression level using RT-PCR technique. In this study, we sequenced one part of vitellogenin cDNA and measured vitellogenin mRNA induction. For the sequence analysis of vitellogenin DNA, reverse transcription was performed with RNA template prepared from the liver using primers based on the cDNA of fathead minnow. The products were running on the 1% agarose gel electrophoresis and the slices of amplified cDNA fragments were cloned into pGEM-T easy vector. The cDNA clones were applied to the automatic DNA sequencer. The cDNA was found to be 742 bp long, which is a partial sequence of vitellogenin mRNA. When we compared the homology of the deduced amino acids, the partial sequence showed a high degree of identity with those of Cyprinus carpio, Pimephales promelas and Danio reiro, with 94, 84, 81%, respectively. Reverse transcription PCR was also carried out with total RNA isolated from adult crucian carp exposed 17-ethinylestradiol and nonylphenol. Vitellogenin mRNA of adult male crucian carp was significantly induced in the chemical treated fish in a dose-dependent manner.
Key words: crucian carp, vitellogenin, cDNA, nonylphenol
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