Bioluminescence of Aequorea macrodactyla, a Common Jellyfish Species in the East China Sea

Abstract:

Studies of the bioluminescent mechanisms of jellyfish have been mainly confined to one species, Aequorea victoria. We describe the luminescent system of another species, Aequorea macrodactyla, which is commonly found in the warmer waters on the coastal region of East China Sea. The luminescent system of this species consists of a green fluorescent protein (GFP) and one or more aequorins. The GFP gene is 1042 bp. It encompasses a coding sequence of 717 bp organized as 3 exons, and it is predicted to specify a 27-kDa peptide, which shares 80% amino acid sequence identity with the GFP of A. victoria. The entire coding sequence was cloned into the pTO-T7 expression vector and expressed in Escherichia coli. Compared with GFP of A. victoria, the purified expressed protein exhibited an excitation peak at a higher wavelength of 476 nm and an emission peak at a lower wavelength of 496 nm, with a higher quantum yield of 1.0. The other photoprotein, aequorin, is encoded in a single open reading frame of 585 bp specifying a 23-kDa apoprotein. The gene was cloned in to the same expression vector and expressed in E. coli. The activity of the photoprotein was reconstituted by incubating the expressed apoprotein with coelenterazine f. In the presence of Ca²⁺ the reconstituted aequorin exhibits an emission peak at 470 nm. The kinetics of regeneration and the photoactivities of the reconstituted aequorins of the 2 species of jellyfish are similar. Nevertheless, Aequorea macrodactyla is expected to appear brighter and more ``blue'' than Aequorea victorea because of the differences in the photoactivity of their GFPs.