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KMID : 1024220110260010005
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Environmental Health and Toxicology : EHT
2011 Volume.26 No. 1 p.5 ~ p.0
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Molecular Cloning and Characterization of the Estrogen Receptor from the Slender Bitterling (Acheilognathus yamatsutae)
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Kim Jong-Geuk
Kim Ha-Ryong
Park Yong-Joo
Chung Kyu-Hyuck
Oh Seung-Min
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Abstract
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Objectives: In order to identify the possibility of slender bitterling (SB) (Acheilognathus yamatsutae) being used as a test species for estrogenic endocrine disrupting chemicals (EEDCs), we carried out the cloning and sequence characterization of the estrogen receptor (ER).
Methods: The ER from a slender bitterling was obtained by reverse transcriptase-polymerase chain reaction (RT-PCR), 5’- and 3’-rapid amplification of cDNA ends (5’-RACE and 3’-RACE) and T-vector cloning. The expression of ER mRNA was also analyzed in six tissues (brain, liver, kidney, gill, gonad, and intestines) by real-time PCR.
Results: We obtained an ER from the slender bitterling. The SB ER cDNA was 2189 base pairs (bp) in length and contained a 1707 bp open reading frame that encoded 568 amino acid residues. The SB ER amino acid sequence clustered in a monophyletic group with the ERα of other fish, and was more closely related to zebrafish ERα (88% identity) than to the ERα of other fish. The SB ER cDNA was divided into A/B, C, D, E and F domains. The SB ER has conserved important sequences for ER functions, such as the DNA binding domain (D domain), which are consistent with those of other teleosts.
Conclusions: The ER of the slender bitterling could provide basic information in toxicological studies of EEDCs in the slender bitterling.
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KEYWORD
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Estrogen receptor, Estrogenic endocrine disrupting effects, Gene cloning, Slender bitterling
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