12th International Symposium on Bioluminescence & Chemiluminescence

Symposium abstracts:

Secreted luciferase of copepode Metridia longa - a novel bioluminescent reporter enzyme

Markova, S.V.¹, Golz, S.², Burakova, L.P.¹, Bondar, V.S.¹, Frank, L.A.¹, Kalthof, B.², Vysotski, E.S.¹

1. Photobiology Lab, Institute of Biophysics RAS, SB, Krasnoyarsk 660036, Russia
2. Bayer AG, PH- R Molecular Screening Technology, 42096 Wuppertal, Germany

Metridia longa is a small crustacean which habitats in the open sea. When disturbed, the animal produces a bright luminous cloud by ejecting a mixture of luciferin and luciferase into the seawater. Aim of this study was the cloning of this unknown luciferase, using an expression cDNA library and a functional screening method. Several related cDNAs encoding coelenterazine-dependent luciferases were identified. Expression of these luciferases in mammalian cells revealed the secretion of luciferase activity into the culture medium. A N-terminal 17 aa residues secretion signal peptide was identified using a neural network trained with signal peptide cleavage sites. The luciferases were expressed in E. coli cells, purified and their biochemical properties were determined. Work was supported by Bayer AG (Germany).

This is a preprint of an article accepted for publication in Luminescence: Copyright 2001 John
Wiley & Sons, Ltd (Wiley website)