Cloning and Expression of the Green Fluorescent Protein from the Clinging Jellyfish, Gonionemus​​ vertens u​sing RNA-Seq.

The clinging jellyfish, ​Gonionemus vertens, ​is a fluorescent hydrozoan native to the Pacific that grows to ~2.5 cm in diameter in the adult medusa stage. ​G. vertens first appeared in the Shrewsbury and Navesink Rivers in New Jersey in June of 2016. In 2018, the organism was found in the Metedeconk River and Tices Shoal in Barnegat Bay, NJ. One remarkable feature of this jellyfish is it’s brilliant green fluorescence when illuminated with blue light (470 nm). It is assumed that this is due to the presence of a green fluorescent protein (GFP) similar to the one first described in another hydrozoan, ​Aequorea victoria. Unlike the ​A. victoria GFP, the ​G. vertens GFP (GvFP) has significantly different excitation and emission spectra (λ​Max = 501.4 ± 0.2 nm and λ​max = 512.9 ± 0.2 nm, respectively) and the shortest known Stokes radius for a GFP (11.5 nm). Since the GvFP has unique biophysical characteristics I propose to clone, sequence and characterize this protein to understand these differences. I will employ amplification by PCR using degenerate primers and NGS transcriptome analysis to find and sequence the gene(s) encoding this important protein. Once isolated I propose to subclone the full-length gene into a plasmid expression vector which should permit unlimited protein production of the recombinant GvFP in ​E. coli​ ​to study its unique biophysical properties​.