Тип публикации: статья из журнала
Год издания: 2012
Идентификатор DOI: 10.1016/j.bbrc.2011.11.063
Ключевые слова: Bioluminescence, Coelenterazine, Mammalian expression, Secretion, luciferase, amino acid sequence, animal cell, article, bacterial cell, controlled study, copepod, enzyme activity, Escherichia coli, gene deletion, human, human cell, Metridia longa, nonhuman, priority journal, protein expression, signal transduction, stomach secretion, Animals, Copepoda, HEK293 Cells, Humans, Luciferases, Molecular Sequence Data, Sequence Deletion, Mammalia, 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 Показать полностьюpeptide 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.
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