High-active truncated luciferase of copepod Metridia longa

Тип публикации: статья из журнала

Год издания: 2012

Идентификатор DOI: 10.1016/j.bbrc.2011.11.063

Ключевые слова: Bioluminescence, Coelenterazine, Mammalian expression, Secretion, Bioluminescence, Coelenterazine, Mammalian expression, Secretion, luciferase, amino acid sequence, animal cell, article, bacterial cell, bioluminescence, controlled study, copepod, enzyme activity, Escherichia coli, gene deletion, human, human cell, Metridia longa, nonhuman, priority journal, protein expression, signal transduction, stomach secretion, Amino Acid Sequence, Animals, Copepoda, Escherichia coli, HEK293 Cells, Humans, Luciferases, Molecular Sequence Data, Sequence Deletion, Copepoda, Escherichia coli, Mammalia, Metridia longa, Metridia luciferase

Аннотация: 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. (C) 2011 Elsevier Inc. All rights reserved.

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Издание

Журнал: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS

Выпуск журнала: Vol. 417, Is. 1

Номера страниц: 98-103

ISSN журнала: 0006291X

Место издания: SAN DIEGO

Издатель: ACADEMIC PRESS INC ELSEVIER SCIENCE

Авторы

  • Markova Svetlana V.
  • Burakova Ludmila P.
  • Vysotski Eugene S. (Russian Acad Sci, Photobiol Lab, Inst Biophys, Siberian Branch, Krasnoyarsk 660036, RussiaSiberian Fed Univ, Dept Biophys, Krasnoyarsk 660041, Russia)

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