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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Markova S.V., Burakova L.P., Vysotski E.S.
Заглавие : High-active truncated luciferase of copepod Metridia longa
Колич.характеристики :6 с
Место публикации : Biochem. Biophys. Res. Commun.: ACADEMIC PRESS INC ELSEVIER SCIENCE, 2012. - Vol. 417, Is. 1. - С. 98-103. - ISSN 0006-291X, DOI 10.1016/j.bbrc.2011.11.063
Примечания : Cited References: 31. - This study was supported by the Grants 16.512.11.2141 and 64987.2010.4 of the Ministry of Education and Science of Russian Federation.
Предметные рубрики: COELENTERAZINE-BINDING PROTEIN
REPORTER-GENE-EXPRESSION
RENILLA LUCIFERASE
GAUSSIA LUCIFERASE
LIGHT-EMITTER
IN-VIVO
BIOLUMINESCENCE
PHOTOPROTEINS
CDNA
SUBSTRATE
Ключевые слова (''Своб.индексиров.''): bioluminescence--coelenterazine--mammalian expression--secretion
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 coil 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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