title: High-active truncated luciferase of copepod Metridia longa.
abstract: The technology of real-time imaging in living cells is crucial for understanding of intracellular events. For this purpose, bioluminescent reporters have been introduced as sensitive and convenient tools. Metridia luciferase (MLuc) from the copepod Metridia longa is a coelenterazine-dependent luciferase containing a natural signal peptide for secretion. We report the high-active MLuc mutants with deletion of the N-terminal variable part of amino acid sequence. The MLuc variants were produced in Escherichia coli cells, converted to an active protein, and characterized. We demonstrate that the truncated MLucs have significantly increased bioluminescent activity as against the wild type enzyme but substantially retain other properties. One of the truncated variants of MLuc was transiently expressed in HEK 293 cells. The results clearly suggest that the truncated Metridia luciferase is well suited as a secreted reporter ensuring higher detection sensitivity in comparison with a wild type enzyme.
authors.affiliation: Institute of Biophysics, Siberian Branch, Russian Academy of Sciences, Krasnoyarsk, Russia.
authors.forename: Svetlana V
authors.initials: SV
authors.lastname: Markova
authors.forename: Ludmila P
authors.initials: LP
authors.lastname: Burakova
authors.forename: Eugene S
authors.initials: ES
authors.lastname: Vysotski
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D033342:Copepoda / Q000201:enzymology*; D004926:Escherichia coli / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D017384:Sequence Deletion
chemical_list: D008156:Luciferases