Тип публикации: статья из журнала
Год издания: 2017
Идентификатор DOI: 10.1134/S0006350917030101
Ключевые слова: bioluminescent reporter, cell viability, coelenterazine, luciferase, melanoma
Аннотация: Secreted reporter proteins provide monitoring of intracellular events in real time without cell destruction. To create human melanoma cell lines that enables noninvasive bioluminescent monitoring of metabolic activity, a comparison of the efficiency of isoforms and mutant variants of luciferase from the Metridia longa as secreted reporter proteins in the cells of human melanoma lines Mel IL was conducted. The MLM3 deletion mutant had the highest activity in the medium of two studied isoforms and two deletion mutants of secreted M. longa luciferase during the Mel IL melanoma cell transfection. It was established that optimization of the gene structure of the selected MLM3 variant for expression in human cells increases the level of bioluminescent activity in the Mel IL cells by almost an order of magnitude. A stable Mel IL melanoma cell line with constitutive expression of the humanized hMLM3 reporter gene was obtained and characterized. The linear range of identification of living cells by the hMLM3 reporter activity was more than three orders of magnitude with a sensitivity of detection of 10 cells.
Издание
Журнал: Biophysics
Выпуск журнала: Т. 62, № 3
Номера страниц: 503-507
ISSN журнала: 00063509
Место издания: Москва
Издатель: Pleiades Publishing, Ltd. (Плеадес Паблишинг, Лтд)
Персоны
- Markova S.V. (Siberian Federal University)
- Malikova N.P. (Blokhin Russian Cancer Research Center)
- Vysotski E.S. (Blokhin Russian Cancer Research Center)
- Frank L.A. (Siberian Federal University)
- Gitelson I.I. (Siberian Federal University)
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