Environ Health Toxicol.  2011 ;26(1):e2011005.

Molecular Cloning and Characterization of the Estrogen Receptor from the Slender Bitterling (Acheilognathus yamatsutae)

• Kim JG
• Kim HR
• Park YJ
• Chung KH
• Oh SM

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.

Keyword

Estrogen receptor; Estrogenic endocrine disrupting effects; Gene cloning; Slender bitterling

MeSH Terms

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
DNA
DNA, Complementary
Endocrine Disruptors
Estrogen Receptor alpha
Estrogens
RNA, Messenger