Библиотека института биофизики СО РАН

Главная

Базы данных

Труды сотрудников ИБФ СО РАН - результаты поиска

Вид поиска

Каталог книг и продолжающихся изданий библиотеки Института биофизики СО РАН

Иностранные журналы библиотеки Института биофизики СО РАН

Отечественные журналы библиотеки Института биофизики СО РАН

Каталог диссертаций ИБФ СО РАН

Труды сотрудников ИБФ СО РАН

Область поиска

в найденном

Формат представления найденных документов:
полный	информационный	краткий

Отсортировать найденные документы по:
автору	заглавию	году издания	типу документа

Поисковый запрос: (<.>K=Reporter<.>)

Общее количество найденных документов : 33
Показаны документы с 1 по 20

1-20 21-33

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Borisova V.V., Pyshnaya I.A., Pyshnyi D.V., Frank L.A.
Заглавие : A Highly Sensitive and Rapid Method for the Detection of DNA Fragments Using the Photoprotein Obelin as a Reporter
Колич.характеристики :7 с
Коллективы :
Место публикации : Russ. J. Bioorg. Chem.: MAIK NAUKA/INTERPERIODICA/SPRINGER, 2008. - Vol. 34, Is. 6. - С. 709-715. - ISSN 1068-1620, DOI 10.1134/S1068162008060101
Примечания : Cited References: 13. - This work was supported the program Molecular and Cellular Biology (project no. 10.6), integration grants of the Siberian Division of the Russian Academy of Sciences (73 and 55), CRDF, and the Russian Foundation for Basic Research (project nos. 06-04-49263-a and 06-04-08076-ofi).
Предметные рубрики: BIOLUMINESCENT IMMUNOASSAY
Ключевые слова (''Своб.индексиров.''): obelin--bioluminescent hybridization assay--pcr
Аннотация: The recombinant Ca(2+)-activated photoprotein obelin was used as a reporter protein in a solid-phase bioluminescent hybridization DNA assay. Oligonucleotide probes were immobilized on the surface of polymer methacrylate beads or microbiological plates of different types. A 30-mer oligonucleotide or its derivative with the biotin residue on the 3'-terminus, as well as a denatured double-stranded PCR fragment of the hepatitis C virus with the sequence of the 30-mer oligonucleotide was used as a DNA template. The probe in the hybridization complex was labeled by the elongation of the chain using a Taq DNA polymerase in the presence of biotinylated deoxyuridine triphosphate. The results of the bioluminescent assay were compared with the results of colorimetric analysis obtained with alkaline phosphatase as a reporter protein. It was shown that the use of the bioluminescent obelin label substantially accelerates the DNA detection procedure, provides a high sensitivity of the assay (no less than 10(-15) mol of DNA template), and ensures a quantitative determination of the amount of DNA template in the tested sample.
Найти похожие

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Nemtseva E. V., Gulnov D. V., Gerasimova M. A., Sukovatyi L. A., Burakova L. P., Karuzina N. E., Melnik B. S., Kratasyuk V. A.
Заглавие : Bacterial luciferases from vibrio harveyi and photobacterium leiognathi demonstrate different conformational stability as detected by time-resolved fluorescence spectroscopy
Место публикации : Int. J. Mol. Sci.: MDPI, 2021. - Vol. 22, Is. 19. - Ст.10449. - ISSN 16616596 (ISSN), DOI 10.3390/ijms221910449
Аннотация: Detecting the folding/unfolding pathways of biological macromolecules is one of the urgent problems of molecular biophysics. The unfolding of bacterial luciferase from Vibrio harveyi is well-studied, unlike that of Photobacterium leiognathi, despite the fact that both of them are actively used as a reporter system. The aim of this study was to compare the conformational transitions of these luciferases from two different protein subfamilies during equilibrium unfolding with urea. Intrinsic steady-state and time-resolved fluorescence spectra and circular dichroism spectra were used to determine the stages of the protein unfolding. Molecular dynamics methods were applied to find the differences in the surroundings of tryptophans in both luciferases. We found that the unfolding pathway is the same for the studied luciferases. However, the results obtained indicate more stable tertiary and secondary structures of P. leiognathi luciferase as compared to enzyme from V. harveyi during the last stage of denaturation, including the unfolding of individual subunits. The distinctions in fluorescence of the two proteins are associated with differences in the structure of the C-terminal domain of ?-subunits, which causes different quenching of tryptophan emissions. The time-resolved fluorescence technique proved to be a more effective method for studying protein unfolding than steady-state methods. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.
Scopus
Найти похожие

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Nemtseva, Elena, V, Gulnov, Dmitry, V, Gerasimova, Marina A., Sukovatyi, Lev A., Burakova, Ludmila P., Karuzina, Natalya E., Melnik, Bogdan S., Kratasyuk, Valentina A.
Заглавие : Bacterial Luciferases from Vibrio harveyi and Photobacterium leiognathi Demonstrate Different Conformational Stability as Detected by Time-Resolved Fluorescence Spectroscopy
Колич.характеристики :17 с
Коллективы : Ministry of Science and Higher Education of the Russian Federation [FSRZ-2020-0006]; RFBRRussian Foundation for Basic Research (RFBR) [20-34-90118]; Krasnoyarsk Regional Fund of Science [20-44-243002, 20-44-240006]; RFBRRussian Foundation for Basic Research (RFBR)
Место публикации : Int. J. Mol. Sci.: MDPI, 2021. - Vol. 22, Is. 19. - Ст.10449. - ISSN 1422-0067(eISSN), DOI 10.3390/ijms221910449
Примечания : Cited References:45. - The research was partially funded by the Ministry of Science and Higher Education of the Russian Federation (projects No. FSRZ-2020-0006); by the RFBR and Krasnoyarsk Territory and Krasnoyarsk Regional Fund of Science (projects No. 20-44-243002 and 20-44-240006); and by the RFBR (project No. 20-34-90118).
Предметные рубрики: TRYPTOPHAN FLUORESCENCE
CRYSTAL-STRUCTURE
SUBUNIT
BIOLUMINESCENCE
Аннотация: Detecting the folding/unfolding pathways of biological macromolecules is one of the urgent problems of molecular biophysics. The unfolding of bacterial luciferase from Vibrio harveyi is well-studied, unlike that of Photobacterium leiognathi, despite the fact that both of them are actively used as a reporter system. The aim of this study was to compare the conformational transitions of these luciferases from two different protein subfamilies during equilibrium unfolding with urea. Intrinsic steady-state and time-resolved fluorescence spectra and circular dichroism spectra were used to determine the stages of the protein unfolding. Molecular dynamics methods were applied to find the differences in the surroundings of tryptophans in both luciferases. We found that the unfolding pathway is the same for the studied luciferases. However, the results obtained indicate more stable tertiary and secondary structures of P. leiognathi luciferase as compared to enzyme from V. harveyi during the last stage of denaturation, including the unfolding of individual subunits. The distinctions in fluorescence of the two proteins are associated with differences in the structure of the C-terminal domain of alpha-subunits, which causes different quenching of tryptophan emissions. The time-resolved fluorescence technique proved to be a more effective method for studying protein unfolding than steady-state methods./p
WOS
Найти похожие

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Krasitskaya, Vasilisa V., Chaukina, Valentina V., Abroskina, Maria V., Vorobyeva, Maria A., Ilminskaya, Aleksandra A., Kabilov, Marsel R., Prokopenko, Semyon V., Nevinsky, Georgy A., Venyaminova, Alya G., Frank, Ludmila A.
Заглавие : Bioluminescent aptamer-based sandwich-type assay of anti-myelin basic protein autoantibodies associated with multiple sclerosis
Колич.характеристики :7 с
Коллективы : Russian Foundation for Basic Research (RFBR), Russia [17-315-50027]; Russian State [AAAA-A17-117013050026-4, AAAA-A17-117020210021-7]
Место публикации : Anal. Chim. Acta: ELSEVIER SCIENCE BV, 2019. - Vol. 1064. - С. 112-118. - ISSN 0003-2670, DOI 10.1016/j.aca.2019.03.015. - ISSN 1873-4324(eISSN)
Примечания : Cited References:29. - This work was supported by the Russian Foundation for Basic Research (RFBR), Russia, under the grant No 17-315-50027; Russian State funded budget projects No. AAAA-A17-117013050026-4 and AAAA-A17-117020210021-7.
Предметные рубрики: ANTIBODIES
BIOMARKERS
RNA
Ключевые слова (''Своб.индексиров.''): bioluminescent microassay--rna aptamers--autoantibodies to myelin basic--protein--multiple sclerosis
Аннотация: Bioluminescent solid-phase sandwich-type microassay was developed to detect multiple sclerosis (MS)-associated autoantibodies in human sera. The assay is based on two different 2'-F-Py RNA aptamers against the target autoantibodies as biospecific elements, and Ca2+-regulated photoprotein obelin as a reporter. The paper describes elaboration of the assay and its application to 91 serum samples from patients with clinically definite MS and 86 ones from individuals healthy in terms of MS. Based on the receiver-operator curve (ROC) analysis, the chosen threshold value as clinical decision limit offers sensitivity of 63.7% and specificity of 94.2%. The area under the ROC curve (AUC) value of 0.87 shows a good difference between the groups under investigation. The likelihood ratio of 10.97 proves the diagnostic value of the assay and its potential as one of the laboratory MS-tests. (C) 2019 Elsevier B.V. All rights reserved.
WOS,
Смотреть статью,
Scopus
Найти похожие

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Burakova, Ludmila P., Kudryavtsev, Alexander N., Stepanyuk, Galina A., Baykov, Ivan K., Morozova, Vera V., Tikunova, Nina V., Dubova, Maria A., Lyapustin, Victor N., Yakimenko, Valeri V., Frank, Ludmila A.
Заглавие : Bioluminescent detection probe for tick-borne encephalitis virus immunoassay
Колич.характеристики :7 с
Коллективы : Russian Academy of Sciences, Siberian Branch [139], Russian Academy of Sciences [VI 57.1.1]
Место публикации : Anal. Bioanal. Chem.: SPRINGER HEIDELBERG, 2015. - Vol. 407, Is. 18. - С. 5417-5423. - ISSN 1618-2642, DOI 10.1007/s00216-015-8710-6. - ISSN 1618-2650(eISSN)
Примечания : Cited References:19. - The work was supported by the Russian Academy of Sciences, Siberian Branch, within the framework of the Interdisciplinary Integration Project No. 139 and the State budget allocated to the fundamental research at the Russian Academy of Sciences (project No. VI 57.1.1).
Предметные рубрики: COELENTERAZINE-BINDING PROTEIN
ENZYME-IMMUNOASSAY
RENILLA-MUELLERI
Ключевые слова (''Своб.индексиров.''): tick-borne encephalitis virus--single-chain antibody--luciferase--immunoassay
Аннотация: To facilitate the detection of the tick-borne encephalitis virus (TBEV), the causative agent of one of the most severe human neuroinfections, we have developed an immunoassay based on bioluminescent hybrid protein 14D5a-Rm7 as a detection probe. The protein containing Renilla luciferase as a reporter and a single-chain variable fragment (scFv) of murine immunoglobulin to TBEV as a recognition element was constructed, produced by bacterial expression, purified, and tested. Both domains were shown to reveal their specific biological properties-affinity to the target antigen and bioluminescent activity. Hybrid protein was applied as a label for solid-phase immunoassay of the antigens, associated with the tick-borne encephalitis virus (native glycoprotein E or extracts of the infected strain of lab ticks). The assay demonstrates high sensitivity (0.056 ng of glycoprotein E; 10(4)-10(5) virus particles or 0.1 pg virions) and simplicity and is competitive with conventional methods for detection of TBEV.
WOS,
Scopus
Найти похожие

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Gorokhovatsky A.Y., Shaloiko L.A., Bondar V.S., Vysotski E.S., Maximov E.E., von Doehren H..., Alakhov Y.B.
Заглавие : Cell-free bioluminescent screening of translation inhibitors
Колич.характеристики :5 с
Место публикации : Biotechnol. Appl. Biochem.: PORTLAND PRESS, 1998. - Vol. 27. - С. 259-263. - ISSN 0885-4513
Примечания : Cited References: 21
Предметные рубрики: DIPHTHERIA-TOXIN
MESSENGER-RNA
SYSTEM
PROTEINS
Аннотация: The possibility of creating new screening methods with a cell-free translation system has been demonstrated with a quantitative determination of diphtheria toxin and some antibiotics (puromycin, kanamycin and tetracycline) as examples. The approach proposed follows from the ability of various substances to inhibit protein synthesis. We used a wheat-germ cell-free translation system stabilized by freeze-drying in the presence of trehalose with the mRNA of the Ca2+-activated photoprotein obelin as a reporter template. This freeze-dried cell-free translation system allows prolonged storage of the detecting system before it is required, increases the reproducibility of the results and simplifies the application procedure. The obelin mRNA extends the sensitivity of the method owing to the high sensitivity of detection of the synthesized protein.
Найти похожие

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Markova S.V., Golz S..., Frank L.A., Kalthof B..., Vysotski E.S.
Заглавие : Cloning and expression of cDNA for a luciferase from the marine copepod Metridia longa - A novel secreted bioluminescent reporter enzyme
Колич.характеристики :6 с
Место публикации : J. Biol. Chem.: AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, 2004. - Vol. 279, Is. 5. - С. 3212-3217. - ISSN 0021-9258, DOI 10.1074/jbc.M309639200
Примечания : Cited References: 37
Предметные рубрики: VARGULA-HILGENDORFII LUCIFERASE
GREEN FLUORESCENT PROTEIN
GENE-EXPRESSION
FIREFLY LUCIFERASE
PROMOTER ACTIVITY
MAMMALIAN-CELLS
RECEPTOR
CANCER
PHOTOPROTEINS
LUMINESCENCE
Аннотация: Metridia longa is a marine copepod from which a blue bioluminescence originates as a secretion from epidermal glands in response to various stimuli. We demonstrate that Metridia luciferase is specific for coelenterazine to produce blue light (lambda(max)=480 nm). Using an expression cDNA library and functional screening, we cloned and sequenced the cDNA encoding the Metridia luciferase. The cDNA is an 897-bp fragment with a 656-bp open reading frame, which encodes a 219-amino acid polypeptide with a molecular weight of 23,885. The polypeptide contains an N-terminal signal peptide of 17 amino acid residues for secretion. On expression of the Metridia luciferase gene in mammalian Chinese hamster ovary cells the luciferase is detected in the culture medium confirming the existence of a naturally occurring signal peptide for secretion in the cloned luciferase. The novel secreted luciferase was tested in a practical assay application in which the activity of A2a and NPY2 G-protein-coupled receptors was detected. These results clearly suggest that the secreted Metridia luciferase is well suited as a reporter for monitoring gene expression and, in particular, for the development of novel ultra-high throughput screening technologies.
Найти похожие

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Titushin M.S., Markova S.V., Frank L.A., Malikova N.P., Stepanyuk G.A., Lee J..., Vysotski E.S.
Заглавие : Coelenterazine-binding protein of Renilla muelleri: cDNA cloning, overexpression, and characterization as a substrate of luciferase
Колич.характеристики :8 с
Место публикации : Photochem. Photobiol. Sci.: ROYAL SOC CHEMISTRY, 2008. - Vol. 7, Is. 2. - С. 189-196. - ISSN 1474-905X, DOI 10.1039/b713109g
Примечания : Cited References: 41
Предметные рубрики: CRYSTAL-STRUCTURE
LIGHT-EMISSION
CA2+-REGULATED PHOTOPROTEINS
BIOLUMINESCENT REPORTER
RENIFORMIS LUCIFERASE
ANGSTROM RESOLUTION
RECOMBINANT OBELIN
ENERGY-TRANSFER
EXCITED-STATE
CALCIUM
Аннотация: The Renilla bioluminescent system in vivo is comprised of three proteins-the luciferase, green-fluorescent protein, and coelenterazine-binding protein (CBP), previously called luciferin-binding protein (LBP). This work reports the cloning of the full-size cDNA encoding CBP from soft coral Renilla muelleri, its overexpression and properties of the recombinant protein. The apo-CBP was quantitatively converted to CBP by simple incubation with coelenterazine. The physicochemical properties of this recombinant CBP are determined to be practically the same as those reported for the CBP (LBP) of R. reniformis. CBP is a member of the four-EF-hand Ca2+-binding superfamily of proteins with only three of the EF-hand loops having the Ca2+-binding consensus sequences. There is weak sequence homology with the Ca2+-regulated photoproteins but only as a result of the necessary Ca2+-binding loop structure. In combination with Renilla luciferase, addition of only one Ca2+ is sufficient to release the coelenterazine as a substrate for the luciferase for bioluminescence. This combination of the two proteins generates bioluminescence with higher reaction efficiency than using free coelenterazine alone as the substrate for luciferase. This increased quantum yield, a difference of bioluminescence spectra, and markedly different kinetics, implicate that a CBP-luciferase complex might be involved.
Найти похожие

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Stepanyuk G.A., Unch J..., Malikova N.P., Markova S.V., Lee J..., Vysotski E.S.
Заглавие : Coelenterazine-v ligated to Ca2+-triggered coelenterazine-binding protein is a stable and efficient substrate of the red-shifted mutant of Renilla muelleri luciferase
Колич.характеристики :9 с
Коллективы :
Место публикации : Anal. Bioanal. Chem.: SPRINGER HEIDELBERG, 2010. - Vol. 398, Is. 4. - С. 1809-1817. - ISSN 1618-2642, DOI 10.1007/s00216-010-4106-9
Примечания : Cited References: 39. - This work was supported by grant 09-04-12022 of the Russian Foundation for Basic Research, "Molecular and Cell Biology" program of Russian Academy of Sciences, by the SB RAS grant No. 2, and by the SB RAS Lavrentiev grant for Young Scientists.
Предметные рубрики: GREEN-FLUORESCENT PROTEIN
BIOLUMINESCENT REPORTER
CA2+-REGULATED PHOTOPROTEINS
RENIFORMIS LUCIFERASE
RECOMBINANT OBELIN
GENE-EXPRESSION
IN-VIVO
CDNA
CLONING
PURIFICATION
Ключевые слова (''Своб.индексиров.''): bioluminescence--coelenterazine--calcium--imaging
Аннотация: It has been shown that the coelenterazine analog, coelenterazine-v, is an efficient substrate for a reaction catalyzed by Renilla luciferase. The resulting bioluminescence emission maximum is shifted to a longer wavelength up to 40 nm, which allows the use of some "yellow" Renilla luciferase mutants for in vivo imaging. However, the utility of coelenterazine-v in small-animal imaging has been hampered by its instability in solution and in biological tissues. To overcome this drawback, we ligated coelenterazine-v to Ca2+-triggered coelenterazine-binding protein from Renilla muelleri, which apparently functions in the organism for stabilizing and protecting coelenterazine from oxidation. The coelenterazine-v bound within coelenterazine-binding protein has revealed a greater long-term stability at both 4 and 37 degrees C. In addition, the coelenterazine-binding protein ligated by coelenterazine-v yields twice the total light over free coelenterazine-v as a substrate for the red-shifted R. muelleri luciferase. These findings suggest the possibility for effective application of coelenterazine-v in various in vitro assays.
Найти похожие

10.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Frank, Ludmila A.
Заглавие : Creation of Artificial Luciferases to Expand their Analytical Potential
Колич.характеристики :11 с
Коллективы : RFBR [14-08-00902/14]; [VI 57.1.1]
Место публикации : Comb. Chem. High Throughput Screen: BENTHAM SCIENCE PUBL LTD, 2015. - Vol. 18, Is. 10. - С. 919-929. - ISSN 1386-2073, DOI 10.2174/1386207318666150917100011. - ISSN 1875-5402(eISSN)
Примечания : Cited References:79. - The work was supported by: the RFBR grant No. 14-08-00902/14; the State budget allocated to the fundamental research at the Russian Academy of Sciences (project No. VI 57.1.1).
Предметные рубрики: BIOLUMINESCENT REPORTER APPLICATIONS
COELENTERAZINE-BINDING PROTEIN
Ключевые слова (''Своб.индексиров.''): luciferase--luciferin--photoprotein--bioluminescence--mutagenesis--luciferase-based assay--bioimaging--reporter assay
Аннотация: Bioluminescent proteins have been intensively used as high sensitive reporters in all kinds of binding assays (immuno-, nucleic acid hybridization assays, etc.) and in bioimaging. But natural luciferases do not always meet the requirements set for them as the assay reporters: thermostabitity, definite bioluminescence spectral and kinetics characteristics, stability to chemical modifications, etc. Luciferases with different appropriate characteristics as well as various luciferin derivatives were obtained using mutagenesis and chemical synthesis. Thanks to rigorous efforts of many researchers bioluminescence-based analytical techniques offer a great potential for solving analytical tasks in the field of biotechnology, biomedicine, pharmacology, etc.
WOS
Найти похожие

11.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Frank L. A.
Заглавие : Creation of artificial luciferases to expand their analytical potential
Место публикации : Comb. Chem. High Throughput Screen. - 2015. - Vol. 18, Is. 10. - С. 919-929. - ISSN 13862073 (ISSN)
Ключевые слова (''Своб.индексиров.''): bioimaging--bioluminescence--luciferase--luciferase-based assay--luciferin--mutagenesis--photoprotein--reporter assay
Аннотация: Bioluminescent proteins have been intensively used as high sensitive reporters in all kinds of binding assays (immuno-, nucleic acid hybridization assays, etc.) and in bioimaging. But natural luciferases do not always meet the requirements set for them as the assay reporters: thermostabitity, definite bioluminescence spectral and kinetics characteristics, stability to chemical modifications, etc. Luciferases with different appropriate characteristics as well as various luciferin derivatives were obtained using mutagenesis and chemical synthesis. Thanks to rigorous efforts of many researchers bioluminescencebased analytical techniques offer a great potential for solving analytical tasks in the field of biotechnology, biomedicine, pharmacology, etc. © 2015 Bentham Science Publishers.
Scopus
Найти похожие

12.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Stepanyuk G.A., Xu H..., Wu C.K., Markova S.V., Lee J..., Vysotski E.S., Wang B.C.
Заглавие : Expression, purification and characterization of the secreted luciferase of the copepod Metridia longa from Sf9 insect cells
Колич.характеристики :7 с
Коллективы :
Место публикации : Protein Expr. Purif.: ACADEMIC PRESS INC ELSEVIER SCIENCE, 2008. - Vol. 61, Is. 2. - С. 142-148. - ISSN 1046-5928, DOI 10.1016/j.pep.2008.05.013
Примечания : Cited References: 34. - This work was supported by the National Institutes of Health (Grant 1P50 GM62407), University of Georgia Research Foundation and Georgia Research Alliance, the Russian Foundation for Basic Research and Taiwan National Science Council (Grant 06-0489502) and the program for "Molecular and Cellular Biology" of Russian Academy of Sciences.
Предметные рубрики: VARGULA-HILGENDORFII LUCIFERASE
CRYSTAL-STRUCTURE
RENILLA-RENIFORMIS
GAUSSIA LUCIFERASE
BIOLUMINESCENT REPORTER
OBELIN BIOLUMINESCENCE
ANGSTROM RESOLUTION
MAMMALIAN-CELLS
GENE-EXPRESSION
IN-VIVO
Аннотация: Metridia luciferase is a secreted luciferase from a marine copepod and uses coelenterazine as a substrate to produce a blue bioluminescence This luciferase has been successfully applied as a bioluminescent reporter in mammalian cells. The main advantage of secreted luciferase as a reporter is the capability of measuring intracellular events without destroying the cells or tissues and this property is well suited for development of high throughput screening technologies. However because Metridia luciferase is a Cys-rich protein, Escherichia coli expression systems produce an incorrectly folded protein, hindering its biochemical characterization and application for development of in vitro bioluminescent assays. Here we report the successful expression of Metridia luciferase with its signal peptide for secretion, in insect (Sf9) cells using the baculovirus expression system. Functionally active luciferase secreted by insect cells into the culture media has been efficiently purified with a yield of high purity protein of 2-3mg/L This Metridia luciferase expressed in the insect cell system is a monomeric protein showing 3.5-fold greater bioluminescence activity than luciferase expressed and purified from E. coli. The near coincidence of the experimental mass of Metridia luciferase purified from insect cells with that calculated from amino acid sequence. indicates that luciferase does not undergo post-translational modifications such as phosphorylation or glycosylation and also, the cleavage site of the signal peptide for secretion is at VQA-KS, as predicted from sequence analysis. (c) 2008 Elsevier Inc. All rights reserved.
Найти похожие

13.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Dubovtseva I. Y., 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
Место публикации : Mol Biol (Mosk): NLM (Medline), 2021. - Vol. 55, Is. 4. - С. 667-675. - ISSN 00268984 (ISSN), DOI 10.31857/S0026898421030058
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.
Scopus
Найти похожие

14.

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.
WOS
Найти похожие

15.

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.
Найти похожие

16.

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
Найти похожие

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
Найти похожие

18.

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.
Найти похожие

19.

20.

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).
Найти похожие