Meeting Abstract
Nuclear receptors are transcription factors and include retinoic acid receptor, retinoid X receptor, estrogen and androgen receptors, and the chicken ovalbumin upstream promoter- transcription factor (COUP-TF). This study focuses on the COUP-TF orphan receptor. COUP-TF is involved in development, homeostasis, and reproduction in both vertebrates and invertebrates. The purpose of our study was to clone and sequence the COUP-TF gene from pale sea anemones (Aiptasia pallida), which are increasingly used as model organisms for the study of cnidarian development. Total RNA was extracted from A. pallida anemones and reverse transcribed to produce cDNA. Degenerate primers designed by comparison of the COUP-TF gene sequences from four cnidarian species were used to amplify cDNA by PCR, and PCR products were separated by gel electrophoresis. Bands of the desired length were excised, and amplified cDNA was purified from these bands and then ligated into pGEM-T plasmids. Bacteria were transformed with these recombinant plasmids and cultured overnight. Plasmid DNA was then extracted and sequenced. We successfully cloned and sequenced 929 nucleotides of the A. pallida COUP-TF gene. The DNA-binding and ligand-binding domains of this gene fragment are 77.7 % and 78% homologous to the Acropora millepora coral COUP-TF gene. To our knowledge, COUP-TF is the first nuclear receptor gene to be sequenced from A. pallida.
