Molecular Cloning and Characterization of the Estrogen Receptor from the Slender Bitterling (Acheilognathus yamatsutae).
- Author:
Jong Geuk KIM
1
;
Ha Ryong KIM
;
Yong Joo PARK
;
Kyu Hyuck CHUNG
;
Seung Min OH
Author Information
1. College of Pharmacy, Sungkyunkwan University, Suwon, Korea.
- Publication Type:Original Article
- Keywords:
Estrogen receptor;
Estrogenic endocrine disrupting effects;
Gene cloning;
Slender bitterling
- MeSH:
Amino Acid Sequence;
Animals;
Base Pairing;
Clone Cells;
Cloning, Molecular;
Cloning, Organism;
DNA;
DNA, Complementary;
Endocrine Disruptors;
Estrogen Receptor alpha;
Estrogens;
Gills;
Gonads;
Kidney;
Liver;
Open Reading Frames;
RNA, Messenger;
Zebrafish
- From:Environmental Health and Toxicology
2011;26(1):e2011005-
- CountryRepublic of Korea
- Language:English
-
Abstract:
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 ERalpha of other fish, and was more closely related to zebrafish ERalpha (88% identity) than to the ERalpha 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.
