Cloning and characterization of androgen receptor from bullfrog, Rana catesbeiana

We have cloned and characterized a full-length cDNA of androgen receptor (AR) from the testis of bullfrog, Rana catesbeiana. The cDNA contains an open reading frame of 2328 nucleotides encoding a protein of 776 amino acid residues. The bullfrog AR shows high homology with ARs from other species in its amino acid sequence. Its overall homology with those of African clawed frog, Japanese eel and human is 70, 53, and 63%, respectively. As expected, the N-terminal domain shows much less homology (30-59%) than both DNA-binding domain (85-92%) and ligand binding domain (80-89%). Northern blot analysis detected the bullfrog AR message as a single transcript of around 9kb only in the testis. However, RT-PCR analysis revealed that AR mRNA is also expressed in other tissues although the levels are very low compared to that in the testis. Western blot analysis of whole tissue extracts showed the presence of AR protein in fore brain, heart, and testis. The AR gene is present as a single copy in bullfrog based on Southern blot analysis of genomic DNA. Altogether, the results suggest that the bullfrog AR is evolutionary conserved and may have functions similar to those shown in other species.