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1.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Frank L.A., Borisova V.V., Markova S.V., Malikova N.P., Stepanyuk G.A., Vysotski E.S.
Заглавие : Violet and greenish photoprotein obelin mutants for reporter applications in dual-color assay
Колич.характеристики :6 с
Место публикации : Anal. Bioanal. Chem.: SPRINGER HEIDELBERG, 2008. - Vol. 391, Is. 8. - С. 2891-2896. - ISSN 1618-2642, DOI 10.1007/s00216-008-2223-5
Примечания : Cited References: 22
Предметные рубрики: ANGSTROM RESOLUTION
RECOMBINANT OBELIN
CRYSTAL-STRUCTURE
BIOLUMINESCENCE
AEQUORIN
IMMUNOASSAY
EXPRESSION
CDNA
PURIFICATION
CLONING
Ключевые слова (''Своб.индексиров.''): ca(2+)-regulated photoprotein--bioluminescence--dual-color assay
Аннотация: Two kinds of Ca(2+)-regulated photoprotein obelin with altered color of bioluminescence were obtained by active-center amino acid substitution. The mutant W92F-H22E emits violet light (lambda(max)=390 nm) and the mutant Y139F emits greenish light (lambda (max)=498 nm), with small spectral overlap, both display high activity and stability and thus may be used as reporters. For demonstration, the mutants were applied in dual-color simultaneous immunoassay of two gonadotropic hormones-follicle-stimulating hormone and luteinizing hormone. Bioluminescence of the reporters was simultaneously triggered by single injection of Ca(2+) solution, divided using band-pass optical filters and measured with a two-channel photometer. The sensitivity of simultaneous bioluminescence assay was close to that of a separate radioimmunoassay.
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2.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Markova, Svetlana V., Larionova, Marina D., Burakova, Ludmila P., Vysotski, Eugene S.
Заглавие : The smallest natural high-active luciferase: Cloning and characterization of novel 16.5-kDa luciferase from copepod Metridia longa
Колич.характеристики :6 с
Коллективы : Bayer AG (Germany); Russian Science Foundation [14-14-01119]
Место публикации : Biochem. Biophys. Res. Commun.: ACADEMIC PRESS INC ELSEVIER SCIENCE, 2015. - Vol. 457, Is. 1. - С. 77-82. - ISSN 0006-291X, DOI 10.1016/j.bbrc.2014.12.082. - ISSN 1090-2104(eISSN)
Примечания : Cited References:20. - The cloning of cDNA encoding MLuc7 luciferase of M. longa was supported by Bayer AG (Germany); all other studies - by the grant 14-14-01119 of the Russian Science Foundation. We declare that authors have no conflict of interest.
Предметные рубрики: CDNA CLONING
SECRETED LUCIFERASE
ESCHERICHIA-COLI
EXPRESSION
Ключевые слова (''Своб.индексиров.''): bioluminescence--coelenterazine--copepod luciferase--mammalian--expression--real-time imaging
Аннотация: Coelenterazine-dependent copepod luciferases containing natural signal peptide for secretion are a very convenient analytical tool as they enable monitoring of intracellular events with high sensitivity, without destroying cells or tissues. This property is well suited for application in biomedical research and development of cell-based assays for high throughput screening. We report the cloning of cDNA gene encoding a novel secreted non-allelic 16.5-kDa isoform (MLuc7) of Metridia longa luciferase, which, in fact, is the smallest natural luciferase of known for today. Despite the small size, isoform contains 10 conservative Cys residues suggesting the presence of up to 5 S-S bonds. This hampers the efficient production of functionally active recombinant luciferase in bacterial expression systems. With the use of the baculovirus expression system, we produced substantial amounts of the proper folded MLuc7 luciferase with a yield of similar to 3 mg/L of a high purity protein. We demonstrate that MLuc7 produced in insect cells is highly active and extremely thermostable, and is well suited as a secreted reporter when expressed in mammalian cells ensuring higher sensitivity of detection as compared to another Metridia luciferase isoform (MLuc164) which is widely employed in real-time imaging. (C) 2014 Elsevier Inc. All rights reserved.
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3.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Larionova, Marina D., Markova, Svetlana, V, Tikunova, Nina, V, Vysotski, Eugene S.
Заглавие : The Smallest Isoform ofMetridia longaLuciferase as a Fusion Partner for Hybrid Proteins
Колич.характеристики :16 с
Коллективы : Russian Foundation for Basic ResearchRussian Foundation for Basic Research (RFBR) [18-44-242001]; Krasnoyarsk Regional Fund of Science; ICBFM SB RAS [AAAA-A17-117020210027-9]; Government of Krasnoyarsk Territory
Место публикации : Int. J. Mol. Sci.: MDPI, 2020. - Vol. 21, Is. 14. - Ст.4971. - ISSN 1422-0067(eISSN), DOI 10.3390/ijms21144971
Примечания : Cited References:49. - The reported study was funded by the Russian Foundation for Basic Research (No. 18-44-242001), Government of Krasnoyarsk Territory, Krasnoyarsk Regional Fund of Science (S.V.M, M.D.L., and E.S.V.) and by the Russian State funded budget project of ICBFM SB RAS No. AAAA-A17-117020210027-9 (N.V.T.).
Предметные рубрики: COELENTERAZINE-BINDING PROTEIN
BIOLUMINESCENT REPORTER
GAUSSIA
Аннотация: Bioluminescent proteins are widely used as reporter molecules in various in vitro and in vivo assays. The smallest isoform of Metridia luciferase (MLuc7) is a highly active, naturally secreted enzyme which, along with other luciferase isoforms, is responsible for the bright bioluminescence of marine copepodMetridia longa. In this study, we report the construction of two variants of a hybrid protein consisting of MLuc7 and 14D5a single-chain antibody to the surface glycoprotein E of tick-borne encephalitis virus as a model fusion partner. We demonstrate that, whereas fusion of a single-chain antibody to either N- or C-terminus of MLuc7 does not affect its bioluminescence properties, the binding site on the single-chain antibody influences its binding capacity. The affinity of 14D5a-MLuc7 hybrid protein (K-D= 36.2 nM) where the C-terminus of the single-chain antibody was fused to the N-terminus of MLuc7, appeared to be 2.5-fold higher than that of the reverse, MLuc7-14D5a (K-D= 87.6 nM). The detection limit of 14D5a-MLuc7 hybrid protein was estimated to be 45 pg of the recombinant glycoprotein E. Although the smallest isoform ofM. longaluciferase was tested as a fusion partner only with a single-chain antibody, it is reasonable to suppose that MLuc7 can also be successfully used as a partner for genetic fusion with other proteins.
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4.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Larionova M. D., Markova S. V., Tikunova N. V., Vysotski E. S.
Заглавие : The smallest isoform of Metridia longa luciferase as a fusion partner for hybrid proteins
Место публикации : Int. J. Mol. Sci.: MDPI AG, 2020. - Vol. 21, Is. 14. - Ст.4971. - С. 1-16. - ISSN 16616596 (ISSN), DOI 10.3390/ijms21144971
Аннотация: Bioluminescent proteins are widely used as reporter molecules in various in vitro and in vivo assays. The smallest isoform of Metridia luciferase (MLuc7) is a highly active, naturally secreted enzyme which, along with other luciferase isoforms, is responsible for the bright bioluminescence of marine copepod Metridia longa. In this study, we report the construction of two variants of a hybrid protein consisting of MLuc7 and 14D5a single-chain antibody to the surface glycoprotein E of tick-borne encephalitis virus as a model fusion partner. We demonstrate that, whereas fusion of a single-chain antibody to either N-or C-terminus of MLuc7 does not affect its bioluminescence properties, the binding site on the single-chain antibody influences its binding capacity. The affinity of 14D5a-MLuc7 hybrid protein (KD = 36.2 nM) where the C-terminus of the single-chain antibody was fused to the N-terminus of MLuc7, appeared to be 2.5-fold higher than that of the reverse, MLuc7-14D5a (KD = 87.6 nM). The detection limit of 14D5a-MLuc7 hybrid protein was estimated to be 45 pg of the recombinant glycoprotein E. Although the smallest isoform of M. longa luciferase was tested as a fusion partner only with a single-chain antibody, it is reasonable to suppose that MLuc7 can also be successfully used as a partner for genetic fusion with other proteins. © 2020 by the authors. Licensee MDPI, Basel, Switzerland.
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5.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Markova S.V., Natashin P.V., Vysotski E.S.
Заглавие : The photoprotein obelin as a bioluminescent reporter to monitor protein-protein interactions in vivo and in vitro by protein-fragment complementation assays
Колич.характеристики :1 с
Место публикации : J. Biotechnol.: ELSEVIER SCIENCE BV, 2010. - Vol. 150: 14th International Biotechnology Symposium and Exhibition (IBS-2008) (SEP 14-18, 2010, Rimini, ITALY). - С. S93-S93. - ISSN 0168-1656, DOI 10.1016/j.jbiotec.2010.08.241
Примечания : Cited References: 0
Ключевые слова (''Своб.индексиров.''): bioluminescence--reporter--obelin--protein-protein interactions
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6.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Larionova, Marina D., Markova, Svetlana V., Vysotski, Eugene S.
Заглавие : The novel extremely psychrophilic luciferase from Metridia longa: Properties of a high-purity protein produced in insect cells
Колич.характеристики :7 с
Коллективы : Bayer AG (Germany); Russian Academy of Sciences [01201351504]; RFBR; Government of Krasnoyarsk Territory [16-44-242099]
Место публикации : Biochem. Biophys. Res. Commun.: ACADEMIC PRESS INC ELSEVIER SCIENCE, 2017. - Vol. 483, Is. 1. - С. 772-778. - ISSN 0006-291X, DOI 10.1016/j.bbrc.2016.12.067. - ISSN 1090-2104(eISSN)
Примечания : Cited References:24. - The cloning of cDNAs encoding MLuc2 isoforms of M. longa was supported by Bayer AG (Germany) and the state budget allocated to the fundamental research at the Russian Academy of Sciences (project No. 01201351504); all other studies were funded by RFBR and Government of Krasnoyarsk Territory according to the research project No. 16-44-242099.
Предметные рубрики: CDNA CLONING
EXPRESSION
ENZYME
Ключевые слова (''Своб.индексиров.''): bioluminescence--coelenterazine--bioluminescent reporter--psychrophilic--enzyme--molecular adaptation
Аннотация: The bright bioluminescence of copepod Metridia longa is conditioned by a small secreted coelenterazinedependent luciferase (MLuc). To date, three isoforms of MLuc differing in length, sequences, and some properties were cloned and successfully applied as high sensitive bioluminescent reporters. In this work, we report cloning of a novel group of genes from M. longa encoding extremely psychrophilic isoforms of MLuc (MLuc2-type). The novel isoforms share only similar to 54-64% of protein sequence identity with the previously cloned isoforms and, consequently, are the product of a separate group of paralogous genes. The MLuc2 isoform with consensus sequence was produced as a natively folded protein using baculovirus/ insect cell expression system, purified, and characterized. The MLuc2 displays a very high bioluminescent activity and high thermostability similar to those of the previously characterized M. longa luciferase isoform MLuc7. However, in contrast to MLuc7 revealing the highest activity at 12-17 degrees C and 0.5 M NaCl, the bioluminescence optima of MLuc2 isoforms are at similar to 5 degrees C and 1 M NaCl. The MLuc(2) adaptation to cold is also accompanied by decrease of melting temperature and affinity to substrate suggesting a more conformational flexibility of a protein structure. The luciferase isoforms with different temperature optima may provide adaptability of the M. longa bioluminescence to the changes of water temperature during diurnal vertical migrations. (C) 2016 Elsevier Inc. All rights reserved.
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7.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Krasitskaya, Vasilisa V., Goncharova, Natalia S., Biriukov, Vladislav V., Bashmakova, Eugenia E., Kabilov, Marsel R., Baykov, Ivan K., Sokolov, Aleksey E., Frank, Ludmila A.
Заглавие : The Ca2+-Regulated Photoprotein Obelin as a Tool for SELEX Monitoring and DNA Aptamer Affinity Evaluation
Колич.характеристики :6 с
Коллективы : Russian Foundation for Basic Research (RFBR)Russian Foundation for Basic Research (RFBR) [18-38-00531]
Место публикации : Photochem. Photobiol.: WILEY, 2020. - Article in press. - ISSN 0031-8655, DOI 10.1111/php.13274. - ISSN 1751-1097(eISSN)
Примечания : Cited References:25. - This work has been supported by the Russian Foundation for Basic Research (RFBR) under the grant no 18-38-00531.
Предметные рубрики: CARDIAC TROPONIN-I
BIOLUMINESCENCE
IMMUNOASSAY
APTASENSOR
DIAGNOSIS
Аннотация: Bioluminescent solid-phase analysis was proposed to monitor the selection process and to determine binding characteristics of the aptamer-target complexes during design and development of the specific aptamers. The assay involves Ca2+-regulated photoprotein obelin as a simple, sensitive and fast reporter. Applicability and the prospects of the approach were exemplified by identification of DNA aptamers to cardiac troponin I, a highly specific early biomarker for acute myocardial infarction. Two structurally different aptamers specific to various epitopes of troponin I were obtained and then tested in a model bioluminescent assay.
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8.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Deeva, Anna A., Temlyakova, Evgenia A., Sorokin, Anatoly A., Nemtseva, Elena V., Kratasyuk, Valentina A.
Заглавие : Structural distinctions of fast and slow bacterial luciferases revealed by phylogenetic analysis
Колич.характеристики :5 с
Коллективы : RFBR [16-34-00746 mol_a]; Ministry of Education and Science of the Russian Federation [1762]; state budget allocated to the fundamental research at Russian Academy of Sciences [01201351504]
Место публикации : Bioinformatics: OXFORD UNIV PRESS, 2016. - Vol. 32, Is. 20. - С. 3053-3057. - ISSN 1367-4803, DOI 10.1093/bioinformatics/btw386. - ISSN 1460-2059(eISSN)
Примечания : Cited References:31. - The reported study was partially funded by RFBR according to the research project No. 16-34-00746 mol_a; by the Ministry of Education and Science of the Russian Federation [project No 1762] and by the state budget allocated to the fundamental research at the Russian Academy of Sciences [project No 01201351504].
Аннотация: Motivation: Bacterial luciferases are heterodimeric enzymes that catalyze a chemical reaction, so called bioluminescence, which causes light emission in bacteria. Bioluminescence is vastly used as a reporter system in research tools and commercial developments. However, the details of the mechanisms that stabilize and transform the reaction intermediates as well as differences in the enzymatic kinetics amongst different bacterial luciferases remain to be elucidated. Results: Amino acid sequences alignments for 21 bacterial luciferases (both alpha- and beta-subunits) were analyzed. For alpha-subunit, containing the enzyme active center, 48 polymorphic amino acid positions were identified. According to them, the sequences fell into two distinct groups known as slow and fast based on the decay rate of the bioluminescence reaction. The differences in the enzyme active site induced by structural polymorphism are analyzed.
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9.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Markova, Svetlana V., Larionova, Marina D., Vysotski, Eugene S.
Заглавие : Shining Light on the Secreted Luciferases of Marine Copepods: Current Knowledge and Applications.
Место публикации : Photochemistry and photobiology. - 2018. - ISSN 1751-1097, DOI 10.1111/php.13077
Аннотация: Copepod luciferases - a family of small secretory proteins of 18.4-24.3 kDa, including a signal peptide, are responsible for bright secreted bioluminescence of some marine copepods. The copepod luciferases use coelenterazine as a substrate to produce blue light in a simple oxidation reaction without any additional cofactors. They do not share sequence or structural similarity with other identified bioluminescent proteins including coelenterazine-dependent Renilla and Oplophorus luciferases. The small size, strong luminescence activity and high stability, including thermostability, make secreted copepod luciferases very attractive candidates as reporter proteins which are particularly useful for nondisruptive reporter assays and for high-throughput format. The most known and extensively investigated representatives of this family are the first cloned GpLuc and MLuc luciferases from copepods Gaussia princeps and Metridia longa, respectively. Immediately after cloning these homologous luciferases were successfully applied as bioluminescent reporters in vivo and in vitro, and since then the scope of their applications continues to grow. This review is an attempt to systemize and critically evaluate the data scattered through numerous articles regarding the main structural features of copepod luciferases, their luminescent and physicochemical properties. We also review the main trends of their application as bioluminescent reporters in cell and molecular biology. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
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10.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Davydova, Anna, Krasitskaya, Vasilisa, Vorobjev, Pavel, Timoshenko, Valentina, Tupikin, Alexey, Kabilov, Marsel, Frank, Ludmila, Venyaminova, Alya, Vorobyeva, Mariya
Заглавие : Reporter-recruiting bifunctional aptasensor for bioluminescent analytical assays
Колич.характеристики :7 с
Коллективы : Russian Science FoundationRussian Science Foundation (RSF) [16-14-10296]; Russian State funded budget projects [AAAA-A17-117020210021-7, AAAA-A19-119031890015-0]
Место публикации : RSC Adv.: ROYAL SOC CHEMISTRY, 2020. - Vol. 10, Is. 54. - С. 32393-32399. - ISSN 2046-2069(eISSN), DOI 10.1039/d0ra05117a
Примечания : Cited References:33. - The work was supported by the Russian Science Foundation (grant #16-14-10296), Russian State funded budget projects #AAAA-A17-117020210021-7 to ICBFM SB RAS and #AAAA-A19-119031890015-0 to IBP SB RAS.
Предметные рубрики: DNA APTAMER
RNA APTAMER
OBELIN
PURIFICATION
EXPRESSION
SEQUENCES
Аннотация: We report a novel bioluminescent aptasensor, which consists of 2 '-F-RNA aptamer modules joined into a bi-specific aptamer construct. One aptamer module binds the analyte, then after structural rearrangement the second module recruits non-covalently Ca2+-dependent photoprotein obelin from the solution, thus providing a bioluminescent signal. This concept allows using free protein as a reporter, which brings such advantages as no need for aptamer-protein conjugation, a possibility of thermal re-folding of aptamer component with no harm to a protein, and simpler detection protocol. We developed the new 2 '-F-RNA aptamer for obelin, and proposed the strategy for engineering structure-switching bi-modular aptamer constructs which bind the analyte and the obelin in a sequential manner. With the use of hemoglobin as a model analyte, we showed the feasibility of utilizing the aptasensor in a fast and straightforward bioluminescent microplate assay. With a proper design of a secondary structure, this strategy of aptasensor engineering might be further extended to bi-specific aptamer-based bioluminescent sensors for other analytes of interest.
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11.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Borisova V.V., Frank L.A., Markova S.V., Burakova L.P., Vysotski E.S.
Заглавие : Recombinant Metridia luciferase isoforms: expression, refolding and applicability for in vitro assay
Колич.характеристики :7 с
Коллективы :
Место публикации : Photochem. Photobiol. Sci.: ROYAL SOC CHEMISTRY, 2008. - Vol. 7, Is. 9. - С. 1025-1031. - ISSN 1474-905X, DOI 10.1039/b807271j
Примечания : Cited References: 19. - The work was supported by Bayer AG, by the Russian Foundation for Basic Research grants 05-04-48271 and 06-04-08076, by the joint grant 06-04-89502 of the Russian Foundation for Basic Research and Taiwan National Science Council, and by the "Molecular and Cellular Biology" program of the Russian Academy of Sciences.
Предметные рубрики: BIOLUMINESCENT REPORTER
GAUSSIA LUCIFERASE
CDNA
PROTEINS
CLONING
OVEREXPRESSION
PURIFICATION
MUTAGENESIS
ENZYME
OBELIN
Аннотация: The recombinant coelenterazine-dependent luciferases (isoforms MLuc 164 and MLuc39) from the marine copepod Metridia longa were expressed as inclusion bodies in E. coli cells, dissolved in 6 M guanidinium chloride and folded in conditions developed for proteins containing intramolecular disulfide bonds. One of them (MLuc09) was obtained in an active monomeric form with a high yield. The luciferase bioluminescence is found to be initiated not only by free coelenterazine, but also by Ca2+-dependent coelenterazine-binding protein (CBP) of Renilla muelleri on Ca2+ addition. The use of CBP as a "substrate" provides higher light emission and simultaneously the lower level of background. The high purity MLuc39 can be detected down to attomol with a linear range extending over 5 orders of magnitude. The MLuc39 reveals also a high stability towards heating and chemical modification; the chemically synthesized biotinylated derivatives of the luciferase preserve 35-40% of the initial activity The luciferase applicability as an in vitro bioluminescent reporter is demonstrated in model tandem bioluminescent solid-phase microassay combining the Ca2+-regulated photoprotein obelin and the Metridia luciferase.
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12.

Вид документа : Статья из сборника (однотомник)
Шифр издания :
Автор(ы) : Illarionov B.A., Matveev S.V., Bondar V.S., Skosyrev V.A., Alakhov Y.B.
Заглавие : Obelin as a carrier protein and reporter enzyme for in vitro synthesized small bioactive polypeptides: New approach to obtain sarcotoxin
Колич.характеристики :4 с
Место публикации : BIOLUMINESCENCE AND CHEMILUMINESCENCE: MOLECULAR REPORTING WITH PHOTONS: JOHN WILEY & SONS LTD, 1997. - 9th International Symposium on Bioluminescence and Chemiluminescence (OCT , 1996, WOODS HOLE, MA). - С. 435-438. - ISBN 0-471-97502-8
Примечания : Cited References: 0
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13.

Вид документа : Статья из сборника (однотомник)
Шифр издания :
Автор(ы) : Illarionov B.A., Matveev S.V., Bondar V.S., Skosyrev V.A., Alakhov Y.B.
Заглавие : Obelin as a carrier protein and reporter enzyme for in vitro synthesized small bioactive polypeptides: New approach to obtain sarcotoxin
Колич.характеристики :4 с
Место публикации : BIOLUMINESCENCE AND CHEMILUMINESCENCE: MOLECULAR REPORTING WITH PHOTONS: JOHN WILEY & SONS LTD, 1997. - 9th International Symposium on Bioluminescence and Chemiluminescence (OCT , 1996, WOODS HOLE, MA). - P435-438. - ISBN 0-471-97502-8
Примечания : Cited References: 0
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14.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Lamarcq L.H., Scherer B.J., Phelan M.L., Kalnine N.N., Nguyen Y.H., Kabakova T..., Chen X.Y., Tan M..., Chang C..., Berlon C..., Campos-Gonzalez R..., Gao G.J., Golz S..., Vysotski E.S., Farmer A.A.
Заглавие : Large-scale, high-throughput validation of short hairpin RNA sequences for RNA interference
Колич.характеристики :11 с
Место публикации : J. Biomol. Screen: SAGE PUBLICATIONS INC, 2006. - Vol. 11, Is. 3. - С. 236-246. - ISSN 1087-0571, DOI 10.1177/1087057105284342
Примечания : Cited References: 50
Предметные рубрики: ENZYME FRAGMENT COMPLEMENTATION
ORFEOME VERSION 1.1
SMALL NUCLEAR-RNA
MAMMALIAN-CELLS
GENE-EXPRESSION
SIRNA SEQUENCES
POLYMERASE-III
SELECTION
TRANSCRIPTION
MICROARRAYS
Ключевые слова (''Своб.индексиров.''): rnai--high-throughput screening--target validation--shrna--reporter assay
Аннотация: (shRNAs) is described. Using this approach, 464 shRNAs auainst 116 different genes were screened for knockdown efficacy, enabling rapid identification of effective shRNAS against 74 genes. Statistical analysis of the effects of various criteria on the activity of the shRNAs confirmed that some of the rules thought to govern small interfering RNA (siRNA) activity also apply to shRNAs. These include moderate GC content, absence of internal hairpins, and asymmetric thermal stability. However, the authors did not find strong support for position-specific rules. In addition, analysis of the data suggests that not all genes are equally susceptible to RNA interference (RNAi).
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15.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Stepanyuk G.A., Golz S..., Markova S.V., Frank L.A., Lee J..., Vysotski E.S.
Заглавие : Interchange of aequorin and obelin bioluminescence color is determined by substitution of one active site residue of each photoprotein
Колич.характеристики :7 с
Место публикации : FEBS Lett.: ELSEVIER SCIENCE BV, 2005. - Vol. 579, Is. 5. - С. 1008-1014. - ISSN 0014-5793, DOI 10.1016/j.febslet.2005.01.004
Примечания : Cited References: 49
Предметные рубрики: FIREFLY LUCIFERASE
SEQUENCE-ANALYSIS
CA2+-REGULATED PHOTOPROTEINS
CA2+-DISCHARGED PHOTOPROTEIN
VIOLET BIOLUMINESCENCE
INTRACELLULAR CALCIUM
ENDOPLASMIC-RETICULUM
ANGSTROM RESOLUTION
CRYSTAL-STRUCTURE
APOAEQUORIN CDNA
Ключевые слова (''Своб.индексиров.''): coelenterazine--calcium--reporter protein--mammalian expression--fluorescence spectrum
Аннотация: The bioluminescence spectra from the Ca2+-regulated photoproteins aequorin (lambda(max) = 469 nm) and obelin (lambda(max) = 482 nm) differ because aequorin has an H-bond from its Tyr82 to the bound coelenteramide, not present in obelin at the corresponding Phe88. Substitutions of this Phe88 by Tyr, Trp, or His shifted the obelin bioluminescence to shorter wavelength with F88Y having lambda(max) = 453 nm. Removal of the H-bond by the substitution of Y82F in aequorin shifted its bioluminescence to lambda(max) = 501 nm. All mutants were stable with good activity and were expressible in mammalian cells, thereby demonstrating potential for monitoring multiple events in cells using multi-color detection. (C) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
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16.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Stepanyuk G.A., Golz S..., Markova S.V., Frank L.A., Lee J..., Vysotski E.S.
Заглавие : Interchange of aequorin and obelin bioluminescence color is determined by substitution of one active site residue of each photoprotein
Колич.характеристики :7 с
Место публикации : FEBS Lett.: ELSEVIER SCIENCE BV, 2005. - Vol. 579, Is. 5. - С. 1008-1014. - ISSN 0014-5793, DOI 10.1016/j.febslet.2005.01.004
Примечания : Cited References: 49
Предметные рубрики: FIREFLY LUCIFERASE
SEQUENCE-ANALYSIS
CA2+-REGULATED PHOTOPROTEINS
CA2+-DISCHARGED PHOTOPROTEIN
VIOLET BIOLUMINESCENCE
INTRACELLULAR CALCIUM
ENDOPLASMIC-RETICULUM
ANGSTROM RESOLUTION
CRYSTAL-STRUCTURE
APOAEQUORIN CDNA
Ключевые слова (''Своб.индексиров.''): coelenterazine--calcium--reporter protein--mammalian expression--fluorescence spectrum
Аннотация: The bioluminescence spectra from the Ca2+-regulated photoproteins aequorin (lambda(max) = 469 nm) and obelin (lambda(max) = 482 nm) differ because aequorin has an H-bond from its Tyr82 to the bound coelenteramide, not present in obelin at the corresponding Phe88. Substitutions of this Phe88 by Tyr, Trp, or His shifted the obelin bioluminescence to shorter wavelength with F88Y having lambda(max) = 453 nm. Removal of the H-bond by the substitution of Y82F in aequorin shifted its bioluminescence to lambda(max) = 501 nm. All mutants were stable with good activity and were expressible in mammalian cells, thereby demonstrating potential for monitoring multiple events in cells using multi-color detection. (C) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
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17.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Bashmakova E. E., Krasitskaya V. V., Kudryavtsev A. N., Grigorenko V. G., Frank L. A.
Заглавие : Hybrid Minimal Core Streptavidin–Obelin as a Versatile Reporter for Bioluminescence-based Bioassay
Место публикации : Photochem. Photobiol.: Blackwell Publishing Inc., 2017. - Vol. 93, Is. 2. - С. 548-552. - ISSN 00318655 (ISSN) , DOI 10.1111/php.12648
Аннотация: Ca2+-regulated photoprotein obelin was genetically fused with a minimum-sized core streptavidin. Hybrid protein (SAV–OL) was produced by bacterial expression and applied as a specific bioluminescent probe in diverse solid-phase assays. The obtained results clearly demonstrate specific activity of each domain indicating its proper folding with favorable space orientation. SAV–OL has been shown to be a much more sensitive label than the chemical conjugate of a full-length streptavidin with obelin. © 2016 The American Society of Photobiology
Scopus,
Смотреть статью,
WOS
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18.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Eremeeva E.V., Markova S.V., Vysotski E.S.
Заглавие : Highly active BRET-reporter based on yellow mutant of Renilla muelleri luciferase
Колич.характеристики :4 с
Место публикации : Dokl. Biochem. Biophys.: MAIK NAUKA/INTERPERIODICA/SPRINGER, 2013. - Vol. 450, Is. 1. - С. 147-150. - ISSN 1607-6729, DOI 10.1134/S1607672913030095
Примечания : Cited References: 14. - This work was supported by the Ministry of Education and Science of the Russian Federation (Government Contract no. 16.512.11.2141) and Council of the President of the Russian Federation on Grants and State Support of Leading Scientific Schools (project no. NSh-64987.2010.4).
Предметные рубрики: GREEN-FLUORESCENT PROTEIN
GENE-EXPRESSION
CDNA
CLONING
BIOLUMINESCENCE
RENIFORMIS
Scopus
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19.

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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20.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Dubovtseva, I. Yu, Aksenenko M. B., Nikolaeva E. D., Averchuk A. S., Moshev, A., V, Savchenko A. A., Markova, S., V, Ruksha T. G.
Заглавие : FOXC1-Mediated Effects of miR-204-5p on Melanoma Cell Proliferation
Колич.характеристики :8 с
Место публикации : Mol. Biol.: PLEIADES PUBLISHING INC, 2021. - Vol. 55, Is. 4. - С. 610-617. - ISSN 0026-8933, DOI 10.1134/S0026893321020199. - ISSN 1608-3245(eISSN)
Примечания : Cited References:24
Предметные рубрики: FOXC1
Аннотация: MicroRNAs epigenetically regulate physiological and pathological processes. Previously, we found that miR-204-5p is expressed at low levels in melanoma cells, and an increase in its level leads to a change in proliferation, migration, and invasion of these cancer cells. Now, using bioinformatics analysis, it has been shown that the target of miR-204-5p is FOXC1 transcription factor, which is implicated in carcinogenesis. Using the luciferase reporter assay, it was found that miR-204-5p suppresses expression of the FOXC1 gene by binding to its 3' non-coding region. Transfection of small interfering RNA (siRNA) targeting FOXC1 into melanoma cells caused a decrease in miR-204-5p levels, which is consistent with the generally accepted concept of feedback regulation of miRNA expression by target genes. According to the results of the MTT test and fluorescence microscopy, the proliferation level of melanoma cells under the influence of siRNA to FOXC1 decreased 72 h after transfection. Changes in the ratio of cells by cell cycle phase were analyzed using flow cytometry. Regulatory relationships between FOXC1 and miR-204-5p, and an inhibitory effect of FOXC1 knockdown on melanoma cell proliferation were revealed. Based on the results, it can be assumed that miR-204-5p regulates proliferation of melanoma cells by affecting FOXC1 expression.
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