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1.


   
    The light-sensitive photoprotein berovin from the bioluminescent ctenophore Beroe abyssicola: a novel type of Ca2+-regulated photoprotein / S. V. Markova [et al.] // FEBS J. - 2012. - Vol. 279, Is. 5. - P856-870, DOI 10.1111/j.1742-4658.2012.08476.x. - Cited References: 63. - The authors thank Natalia Chervyakova from Department of Zoology of Invertebrates of Moscow State University for the photo of the White Sea ctenophore Beroe abyssicola. This work was supported by RFBR grant 09-04-00172, by grant 64987.2010.4, Molecular and Cellular Biology program of RAS, and Bayer Pharma AG (Germany). . - ISSN 1742-464X
РУБ Biochemistry & Molecular Biology
Рубрики:
CALCIUM-ACTIVATED PHOTOPROTEINS
   C-TERMINAL PROLINE

   SEQUENCE-ANALYSIS

   MNEMIOPSIS-SP

   COELENTERAZINE-BINDING

   ANGSTROM RESOLUTION

   RECOMBINANT OBELIN

   CRYSTAL-STRUCTURES

   EXCITED-STATE

   CDNA CLONING

Кл.слова (ненормированные):
bioluminescence -- calcium -- coelenterazine -- luciferase -- mammalian expression
Аннотация: Light-sensitive Ca2+-regulated photoproteins are responsible for the bright bioluminescence of ctenophores. Using functional screening, four full-size cDNA genes encoding the same 208-amino-acid polypeptide were isolated from two independent cDNA libraries prepared from two Beroe abyssicola specimens. Sequence analysis revealed three canonical EF-hand calcium-binding sites characteristic of Ca2+-regulated photoproteins, but a very low degree of sequence identity (2729%) with aequorin-type photoproteins, despite functional similarities. Recombinant berovin was expressed in Escherichia coli cells, purified, converted to active photoprotein and characterized. Active berovin has absorption maxima at 280 and 437 nm. The Ca2+-discharged protein loses visible absorption, but exhibits a new absorption maximum at 335 nm. The berovin bioluminescence is blue (?max = 491 nm) and a change in pH over the range 6.09.5 has no significant effect on the light emission spectrum. By contrast, the fluorescence of Ca2+-discharged protein (?ex = 350 nm) is pH sensitive: at neutral pH the maximum is at 420 nm and at alkaline pH there are two maxima at 410 and 485 nm. Like native ctenophore photoproteins, recombinant berovin is also inactivated by light. The Ca2+ concentrationeffect curve is a sigmoid with a slope on a loglog plot of similar to 2.5. Although this curve for berovin is very similar to those obtained for obelin and aequorin, there are evident distinctions: berovin responds to calcium changes at lower concentrations than jellyfish photoproteins and its Ca2+-independent luminescence is low. Recombinant berovin was successfully expressed in mammalian cells, thereby demonstrating potential for monitoring intracellular calcium.

Держатели документа:
[Vysotski, Eugene S.] Russian Acad Sci, Siberian Branch, Inst Biophys, Photobiol Lab, Krasnoyarsk 660036, Russia
[Markova, Svetlana V.
Burakova, Ludmila P.
Malikova, Natalia P.
Frank, Ludmila A.
Vysotski, Eugene S.] Siberian Fed Univ, Dept Biophys, Krasnoyarsk, Russia
[Golz, Stefan] Bayer Pharma AG, Global Drug Discovery, Wuppertal, Germany
ИБФ СО РАН : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Markova, S.V.; Burakova, L.P.; Golz, S...; Malikova, N.P.; Frank, L.A.; Vysotski, E.S.

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2.


   
    Expression, purification and characterization of the secreted luciferase of the copepod Metridia longa from Sf9 insect cells [Text] / G. A. Stepanyuk [et al.] // Protein Expr. Purif. - 2008. - Vol. 61, Is. 2. - P142-148, DOI 10.1016/j.pep.2008.05.013. - 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. . - ISSN 1046-5928
РУБ Biochemical Research Methods + Biochemistry & Molecular Biology + Biotechnology & Applied Microbiology
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.

Держатели документа:
[Stepanyuk, Galina A.
Markova, Svetlana V.
Vysotski, Eugene S.] Russian Acad Sci, Photobiol Lab, Inst Biophys, Siberian Branch, Krasnoyarsk 660036, Russia
[Stepanyuk, Galina A.
Xu, Hao
Wu, Chia-Kuei
Lee, John
Vysotski, Eugene S.
Wang, Bi-Cheng] Univ Georgia, Dept Biochem & Mol Biol, Athens, GA 30602 USA
ИБФ СО РАН : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Stepanyuk, G.A.; Xu, H...; Wu, C.K.; Markova, S.V.; Lee, J...; Vysotski, E.S.; Wang, B.C.

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3.


   
    Cloning and expression of cDNA for a luciferase from the marine copepod Metridia longa - A novel secreted bioluminescent reporter enzyme [Text] / S. V. Markova [et al.] // J. Biol. Chem. - 2004. - Vol. 279, Is. 5. - P3212-3217, DOI 10.1074/jbc.M309639200. - Cited References: 37 . - ISSN 0021-9258
РУБ Biochemistry & Molecular Biology
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.

Держатели документа:
Russian Acad Sci, Siberian Branch, Inst Biophys, Photobiol Lab, Krasnoyarsk 660036, Russia
Bayer AG, Pharma Res Mol Screening Technol, D-42096 Wuppertal, Germany
ИБФ СО РАН : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Markova, S.V.; Golz, S...; Frank, L.A.; Kalthof, B...; Vysotski, E.S.

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4.


   
    Cell-free bioluminescent screening of translation inhibitors [Text] / A. Y. Gorokhovatsky [et al.] // Biotechnol. Appl. Biochem. - 1998. - Vol. 27. - P259-263. - Cited References: 21 . - ISSN 0885-4513
РУБ Biochemistry & Molecular Biology + Biotechnology & Applied Microbiology
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.

Держатели документа:
Russian Acad Sci, Branch Inst Bioorgan Chem, Pushchino 142292, Moscow Region, Russia
RAS, Inst Biophys, Siberian Branch, Krasnoyarsk 660036, Russia
Tech Univ Berlin, Inst Biochem & Mol Biol, D-10587 Berlin, Germany
ИБФ СО РАН : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Gorokhovatsky, A.Y.; Shaloiko, L.A.; Bondar, V.S.; Vysotski, E.S.; Maximov, E.E.; von Doehren, H...; Alakhov, Y.B.

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5.


   
    Large-scale, high-throughput validation of short hairpin RNA sequences for RNA interference [Text] / L. H. Lamarcq [et al.] // J. Biomol. Screen. - 2006. - Vol. 11, Is. 3. - P236-246, DOI 10.1177/1087057105284342. - Cited References: 50 . - ISSN 1087-0571
РУБ Biochemical Research Methods + Biotechnology & Applied Microbiology + Chemistry, Analytical
Рубрики:
ENZYME FRAGMENT COMPLEMENTATION
   ORFEOME VERSION 1.1

   SMALL NUCLEAR-RNA

   MAMMALIAN-CELLS

   GENE-EXPRESSION

   SIRNA SEQUENCES

   POLYMERASE-III

   SELECTION

   TRANSCRIPTION

   MICROARRAYS

Кл.слова (ненормированные):
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).

Держатели документа:
Clontech Labs, Mountain View, CA 94043 USA
BD Biosci Pharmingen, La Jolla, CA USA
Bayer Healthcare AG, Pharma Res, Mol Screening Technol, Wuppertal, Germany
Russian Acad Sci, Inst Biophys, Photobiol Lab, Siberian Branch, Krasnoyarsk, Russia
ИБФ СО РАН : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
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.

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6.


   
    Development of the bioluminescent bioindicators for analyses of pollutions [Text] / V. A. Kratasyuk [et al.] ; ed. J Gottlieb [et al.] // FIELD SCREENING EUROPE : SPRINGER, 1997. - 1st International Conference on Field Screening Europe - Strategies and Techniques for On-Site Investigation and Monitoring of Contaminated Soil, Water and Air (SEP 29-OCT 01, 1997, KARLSRUHE, GERMANY). - P. 207-210. - Cited References: 0 . - ISBN 0-7923-4782-X
РУБ Engineering, Environmental + Environmental Sciences + Geosciences, Multidisciplinary


WOS
Держатели документа:
RAS, SB, Inst Biophys, Krasnoyarsk 660036, Russia
ИБФ СО РАН : 660036, Красноярск, Академгородок, д. 50, стр. 50
Доп.точки доступа:
Kratasyuk, V.A.; Kudryasheva, N.S.; Khendogina, E.V.; Vetrova, E.V.; Kudinova, I.Y.; Gottlieb, J \ed.\; Hotzl, H \ed.\; Huck, K \ed.\

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7.


   
    Biotesting of sewage and river water by liofilyzed luminous bacteria biotest [Text] / A. M. Kuznetsov, E. K. Rodicheva, S. E. Medvedeva ; ed. J Gottlieb [et al.] // FIELD SCREENING EUROPE : SPRINGER, 1997. - 1st International Conference on Field Screening Europe - Strategies and Techniques for On-Site Investigation and Monitoring of Contaminated Soil, Water and Air (SEP 29-OCT 01, 1997, KARLSRUHE, GERMANY). - P. 211-215. - Cited References: 0 . - ISBN 0-7923-4782-X
РУБ Engineering, Environmental + Environmental Sciences + Geosciences, Multidisciplinary


WOS
Держатели документа:
RAS, SB, Inst Biophys, Krasnoyarsk 660036, Russia
ИБФ СО РАН : 660036, Красноярск, Академгородок, д. 50, стр. 50
Доп.точки доступа:
Kuznetsov, A.M.; Rodicheva, E.K.; Medvedeva, S.E.; Gottlieb, J \ed.\; Hotzl, H \ed.\; Huck, K \ed.\

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8.


   
    The smallest natural high-active luciferase: Cloning and characterization of novel 16.5-kDa luciferase from copepod Metridia longa [Text] / S. V. Markova [et al.] // Biochem. Biophys. Res. Commun. - 2015. - Vol. 457, Is. 1. - P77-82, DOI 10.1016/j.bbrc.2014.12.082. - 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. . - ISSN 0006-291X. - ISSN 1090-2104
РУБ Biochemistry & Molecular Biology + Biophysics
Рубрики:
CDNA CLONING
   SECRETED LUCIFERASE

   ESCHERICHIA-COLI

   EXPRESSION

Кл.слова (ненормированные):
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.

WOS
Держатели документа:
Russian Acad Sci, Inst Biophys, Photobiol Lab, Siberian Branch, Krasnoyarsk, Russia.
Siberian Fed Univ, Chair Biophys, Krasnoyarsk, Russia.
ИБФ СО РАН

Доп.точки доступа:
Markova, Svetlana V.; Larionova, Marina D.; Burakova, Ludmila P.; Vysotski, Eugene S.; Bayer AG (Germany); Russian Science Foundation [14-14-01119]

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9.


   
    Simultaneous Genotyping of Four Single Nucleotide Polymorphisms Associated with Risk Factors of Hemostasis Disorders [Text] / E. E. Bashmakova, V. V. Krasitskaya, L. A. Frank // Comb. Chem. High Throughput Screen. - 2015. - Vol. 18, Is. 10. - P930-936, DOI 10.2174/1386207318666150917095903. - Cited References:20. - The study was supported by the grant 14-14-01119 of the Russian Science Foundation. . - ISSN 1386-2073. - ISSN 1875-5402
РУБ Biochemical Research Methods + Chemistry, Applied + Pharmacology &
Рубрики:
ALLELE-SPECIFIC PCR
   FACTOR-V-LEIDEN

   BIOLUMINESCENT IMMUNOASSAY

Кл.слова (ненормированные):
SNP detection -- PEXT reaction -- photoprotein obelin -- bioluminescent -- microassay -- multiplex PCR
Аннотация: Multiplex simultaneous genotyping technique was developed for four polymorphisms in genes coding for blood coagulation factors and homocysteine metabolism which are considered as thrombophilia related mutations: FV Leiden, FII G20210A, MTHFR C677T, and FVII G10976A. It is based on primer extension reaction with the following bioluminescent solid-phase microassay. At that, two different in bioluminescence obelin mutants were applied to simultaneous detection of two gene allelic variants. The assay is carried out in microtiter plate format and provides fast and reliable genotyping of four single nucleotide polymorphisms in four different genes within 2.5 hours. A large number of clinical samples were analyzed and the obtained results were found to be in complete correlation with those obtained by using conventional RT-PCR techniques.

WOS
Держатели документа:
Russian Acad Sci, Siberian Branch, Inst Biophys, Photobiol Lab, Krasnoyarsk 660036, Russia.
Siberian Fed Univ, Krasnoyarsk 660041, Russia.

Доп.точки доступа:
Bashmakova, Eugenia E.; Krasitskaya, Vasilisa V.; Frank, Ludmila A.; Russian Science Foundation [14-14-01119]

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10.


   
    Applications of Luminous Bacteria Enzymes in Toxicology [Text] / V. A. Kratasyuk, E. N. Esimbekova // Comb. Chem. High Throughput Screen. - 2015. - Vol. 18, Is. 10. - P952-959, DOI 10.2174/1386207318666150917100257. - Cited References:88. - The research was supported by the Russian Science Foundation, project No. 15-19-10041. . - ISSN 1386-2073. - ISSN 1875-5402
РУБ Biochemical Research Methods + Chemistry, Applied + Pharmacology &
Рубрики:
NADHFMN-OXIDOREDUCTASE-LUCIFERASE
   HUMIC SUBSTANCES

   BIOLUMINESCENT

Кл.слова (ненормированные):
Bioluminescence -- bioluminescent toxicity enzymatic assay -- immobilization -- of enzymes -- luciferase -- total toxicity
Аннотация: This review describes the principle and applications of bioluminescent enzymatic toxicity bioassays. This type of assays uses bacterial coupled enzyme systems: NADH: FMN-oxidoreductase and luciferase to replace living organisms in developing cost-competitive biosensors for environmental, medical and industrial applications. These biosensors instantly signal chemical and biological hazards and allow for detecting a great amount of toxic compounds with advantages associated with fast results, high sensitivity, simplicity, low cost and safety of the procedure.

WOS
Держатели документа:
Siberian Fed Univ, Krasnoyarsk 660041, Russia.
Russian Acad Sci, Inst Biophys, Siberian Branch, Photobiol Lab, Krasnoyarsk 660036, Russia.

Доп.точки доступа:
Kratasyuk, Valentina A.; Esimbekova, Elena N.; Russian Science Foundation [15-19-10041]

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11.


   
    Creation of Artificial Luciferases to Expand their Analytical Potential [Text] / L. A. Frank // Comb. Chem. High Throughput Screen. - 2015. - Vol. 18, Is. 10. - P919-929, DOI 10.2174/1386207318666150917100011. - 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). . - ISSN 1386-2073. - ISSN 1875-5402
РУБ Biochemical Research Methods + Chemistry, Applied + Pharmacology &
Рубрики:
BIOLUMINESCENT REPORTER APPLICATIONS
   COELENTERAZINE-BINDING PROTEIN

Кл.слова (ненормированные):
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
Держатели документа:
Russian Acad Sci, Siberian Branch, Inst Biophys, Photobiol Lab, Krasnoyarsk 660036, Russia.
Siberian Fed Univ, Krasnoyarsk 660041, Russia.

Доп.точки доступа:
Frank, Ludmila A.; RFBR [14-08-00902/14]; [VI 57.1.1]

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12.


   
    Recent Trends and Advancements in Bioassay Based on Bioluminescent Technologies [Text] / L. A. Frank // Comb. Chem. High Throughput Screen. - 2015. - Vol. 18, Is. 10. - P918-918, DOI 10.2174/138620731810151103112033. - Cited References:0 . - ISSN 1386-2073. - ISSN 1875-5402
РУБ Biochemical Research Methods + Chemistry, Applied + Pharmacology &


WOS
Держатели документа:
Russian Acad Sci, Inst Biophys, Siberian Branch, Krasnoyarsk 660036, Russia.
Доп.точки доступа:
Frank, Ludmila A.

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13.


   
    Recent trends and advancements in bioassay based on bioluminescent technologies / L. A. Frank // Comb. Chem. High Throughput Screen. - 2015. - Vol. 18, Is. 10. - P918 . - ISSN 1386-2073

Scopus
Держатели документа:
Institute of Biophysics, Russian Academy of Sciences, Siberian Branch 50/50, Krasnoyarsk, Russian Federation

Доп.точки доступа:
Frank, L. A.

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14.


   
    Creation of artificial luciferases to expand their analytical potential / L. A. Frank // Comb. Chem. High Throughput Screen. - 2015. - Vol. 18, Is. 10. - P919-929 . - ISSN 1386-2073
Кл.слова (ненормированные):
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
Держатели документа:
Photobiology Laboratory, Institute of Biophysics, Russian Academy of Sciences, Siberian Branch, Krasnoyarsk, Russian Federation
Siberian Federal University, Svobodnii ave., 79, Krasnoyarsk, Russian Federation

Доп.точки доступа:
Frank, L. A.

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15.


   
    Simultaneous genotyping of four single nucleotide polymorphisms associated with risk factors of hemostasis disorders / E. E. Bashmakova, V. V. Krasitskaya, L. A. Frank // Comb. Chem. High Throughput Screen. - 2015. - Vol. 18, Is. 10. - P930-936 . - ISSN 1386-2073
Кл.слова (ненормированные):
Bioluminescent microassay -- Multiplex PCR -- PEXT reaction -- Photoprotein obelin -- SNP detection
Аннотация: Multiplex simultaneous genotyping technique was developed for four polymorphisms in genes coding for blood coagulation factors and homocysteine metabolism which are considered as thrombophilia related mutations: FV Leiden, FII G20210A, MTHFR C677T, and FVII G10976A. It is based on primer extension reaction with the following bioluminescent solid-phase microassay. At that, two different in bioluminescence obelin mutants were applied to simultaneous detection of two gene allelic variants. The assay is carried out in microtiter plate format and provides fast and reliable genotyping of four single nucleotide polymorphisms in four different genes within 2.5 hours. A large number of clinical samples were analyzed and the obtained results were found to be in complete correlation with those obtained by using conventional RT-PCR techniques. © 2015 Bentham Science Publishers.

Scopus
Держатели документа:
Photobiology Laboratory, Institute of Biophysics, Russian Academy of Sciences, Siberian Branch, Krasnoyarsk, Russian Federation
Siberian Federal University, Svobodnii Ave., 79, Krasnoyarsk, Russian Federation

Доп.точки доступа:
Bashmakova, E. E.; Krasitskaya, V. V.; Frank, L. A.

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16.


   
    Applications of luminous bacteria enzymes in toxicology / V. A. Kratasyuk, E. N. Esimbekova // Comb. Chem. High Throughput Screen. - 2015. - Vol. 18, Is. 10. - P952-959 . - ISSN 1386-2073
Кл.слова (ненормированные):
Bioluminescence -- Bioluminescent toxicity enzymatic assay -- Immobilization of enzymes -- Luciferase -- Total toxicity
Аннотация: This review describes the principle and applications of bioluminescent enzymatic toxicity bioassays. This type of assays uses bacterial coupled enzyme systems: NADH:FMN-oxidoreductase and luciferase to replace living organisms in developing cost-competitive biosensors for environmental, medical and industrial applications. These biosensors instantly signal chemical and biological hazards and allow for detecting a great amount of toxic compounds with advantages associated with fast results, high sensitivity, simplicity, low cost and safety of the procedure. © 2015 Bentham Science Publishers.

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Держатели документа:
Siberian Federal University, Svobodnii Ave., 79, Krasnoyarsk, Russian Federation
Photobiology Laboratory, Russian Academy of Sciences, Siberian Branch, Institute of Biophysics SB RAS, Akademgorodok 50/50, Krasnoyarsk, Russian Federation

Доп.точки доступа:
Kratasyuk, V. A.; Esimbekova, E. N.

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17.


   
    Concentrations of metals in water of the Yenisei River between Krasnoyarsk and the Angara River outfall in 2010-2015 / D. V. Dementyev [и др.] // Bull. Tomsk Polytech. Univ. Geo Assets Eng. - 2017. - Vol. 328, Is. 3. - С. 54-63 . - ISSN 2500-1019
Кл.слова (ненормированные):
Heavy metals -- Microelements -- Screening -- The Yenisei River -- Water -- Water quality
Аннотация: The relevance of the study is determined by the necessity to obtain the data on concentrations of chemical elements in water of the Yenisei River, which is continuously affected by human activities. The aim of the study is to determine concentrations of such chemical elements as Na, K, Ca, Mg, Zn, Cu, Ba, Al, Mn, U, Mo, Cr, Ni, As, Co, Sr, Fe, Pb, Cd, and Bi in water of the Yenisei River between the city of Krasnoyarsk and the outfall of the Angara River. The methods used in the study. Total contents of the elements were determined by inductively coupled plasma mass spectrometry, using an Agilent 7500a instrument. The results. The study showed the changes in average concentrations of 20 chemical elements in water of the Yenisei River between the city of Krasnoyarsk and the outfall of the Angara River (the village Strelka), for 2010-2015. Comparison of the data obtained with the levels of maximum permissible concentrations showed that the examined part of the Yenisei River contained the increased concentrations of Fe, Cu, Mn, Al, and Zn. For the study period, the magnitudes of average concentrations of Fe reached 1...2 maximum permissible concentrations (MPC) and Cu concentrations reached 1...5 MPC The average concentrations of Mn amounted to 1...3 MPC in the parts of the river over 90 km downstream of Krasnoyarsk (at the villages Atamanovo and Strelka). Cu, Fe, and Zn concentrations tend to increase somewhat in the parts of the river over 90 km downstream of Krasnoyarsk, which may be caused by the influence of the industrial complex of the town Zheleznogorsk and the outfall of the Kan River, with the large industrial facilities located along its banks, in Zelenogorsk, in particular.

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Держатели документа:
Institute of Biophysics, Siberian Branch, Russian Academy of Sciences FRC KSC SB RAS, 50/50 Akademgorodok, Krasnoyarsk, Russian Federation
ICCT SB RAS FRC KSC SB RAS, 50/24 Akademgorodok, Krasnoyarsk, Russian Federation

Доп.точки доступа:
Dementyev, D. V.; Bolsunovsky, A. Y.; Borisov, R. V.; Alexandrova, Y. V.

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18.


   
    The disulfide-rich Metridia luciferase refolded from E. coli inclusion bodies reveals the properties of a native folded enzyme produced in insect cells / S. V. Markova [et al.] // J. Photochem. Photobiol. B-Biol. - 2017. - Vol. 175. - P51-57, DOI 10.1016/j.jphotobiol.2017.08.024. - Cited References:30. - These studies were funded by RFBR and the Government of Krasnoyarsk Territory according to the research project No. 16-44-242099 and the state budget allocated to the fundamental research at the Russian Academy of Sciences (project No. 0356-2016-0712). . - ISSN 1011-1344
РУБ Biochemistry & Molecular Biology + Biophysics
Рубрики:
GAUSSIA-PRINCEPS LUCIFERASE
   ESCHERICHIA-COLI

   EXPRESSION

   PROTEIN

Кл.слова (ненормированные):
Copepod luciferase -- Disulfide bonds -- Cysteine-rich protein -- Oxidative -- refolding
Аннотация: The bioluminescence of a marine copepod Metridia Tonga is determined by a small secreted coelenterazine-dependent luciferase that uses coelenterazine as a substrate of enzymatic reaction to generate light (lambda(max) = 480 nm). To date, four different isoforms of the luciferase differing in size, sequences, and properties have been cloned by functional screening. All of them contain ten conserved Cys residues that suggests up to five S-S intramolecular bonds per luciferase molecule. Whereas the use of copepod luciferases as bioluminescent reporters in biomedical research in vivo is growing from year to year, their application for in vitro assays is still limited by the difficulty in obtaining significant amounts of luciferase. The most cost-effective host for producing recombinant proteins is Escherichia coli. However, prokaryotic and eukaryotic cells maintain the reductive environment in cytoplasm that hinders the disulfide bond formation and consequently the proper folding of luciferase. Here we report the expression of the MLuc7 isoform of M. longa luciferase in E. colt cells and the efficient procedure for refolding from inclusion bodies yielding a high-active monomeric protein. Furthermore, in a set of identical experiments we demonstrate that bioluminescent and structural features of MLuc7 produced in bacterial cells are identical to those of MLuc7 isoform produced from culture medium of insect cells. Although the yield of high-purity protein is only 6 mg/L, the application of E. coil cells to produce the luciferase is simpler and more cost-effective than the use of insect cells. We expect that the suggested technology of Metridia luciferase production allows obtaining of sufficient amounts of protein both for the development of novel in vitro analytical assays with the use of MLuc7 as a label and for structural studies.

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Держатели документа:
RAS, Krasnoyarsk Sci Ctr SB, Fed Res Ctr, Photobiol Lab,Inst Biophys SB, Krasnoyarsk, Russia.
Siberian Fed Univ, Krasnoyarsk, Russia.

Доп.точки доступа:
Markova, Svetlana V.; Larionova, Marina D.; Gorbunova, Darya A.; Vysotski, Eugene S.; RFBR; Government of Krasnoyarsk Territory [16-44-242099]; Russian Academy of Sciences [0356-2016-0712]

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19.


   
    Is Body Mass Index a potential biomarker for anemia in obese adolescents? / S. Pande, R. Ranjan, V. A. Kratasyuk // J. Nutr. Intermediary Metab. - 2019. - Vol. 15. - P1-2, DOI 10.1016/j.jnim.2018.11.001 . - ISSN 2352-3859
Кл.слова (ненормированные):
Anemia -- Body Mass Index -- Hepcidin -- Leptin -- Obesity
Аннотация: The two paradoxical major health problems namely obesity and anemia are confirmed to affect millions around the world. Hepcidin, a protein synthesized in liver is a negative iron binding regulator. There is an affirmative relation between hepcidin and leptin levels and an inverse co-relation between hepcidin and iron status due to inflammation mediated by obesity in adolescents. So this implicates an alliance between anemia and obesity wherein weight reduction can be a powerful medium to improve iron absorption in obese adolescents. Also the Body Mass Index can serve as a preliminary non-invasive screening tool to identify potential adolescents prone to anemia. © 2018

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Держатели документа:
Laboratory of Bioluminescent Biotechnologies, Department of Biophysics, Institute of Fundamental Biology and Biotechnology, Siberian Federal University, 79 Svobodny Prospect, Krasnoyarsk, 660041, Russian Federation
Institute of Biophysics SB RAS, Federal Research Center ‘Krasnoyarsk Science Center SB RAS’, Akademgorodok 50/50, Krasnoyarsk, 660036, Russian Federation

Доп.точки доступа:
Pande, S.; Ranjan, R.; Kratasyuk, V. A.

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20.


   
    Screening of biopolymeric materials for cardiovascular surgery toxicity—Evaluation of their surface relief with assessment of morphological aspects of monocyte/macrophage polarization in atherosclerosis patients / N. G. Menzyanova [et al.] // Toxicol. Rep. - 2019. - Vol. 6. - P74-90, DOI 10.1016/j.toxrep.2018.11.009 . - ISSN 2214-7500
Кл.слова (ненормированные):
Atherosclerosis -- Cell morphology -- Intravascular stenting -- Macrophages -- Monocytes -- Polyhydroxyalkanoates
Аннотация: The morphotypes of human macrophages (MPh) were studied in the culture on nano-structured biopolymer substrates, made from polyhydroxyalcanoates (PHAs) of five various monomer compositions, followed by the solvent evaporation. Its surface relief, which was further in direct contact with human cells in vitro, was analyzed by atomic force microscopy (AFM) and scanning electron microscopy (SEM). It was shown, that the features of the micro/nano relief depend on the monomeric composition of the polymer substrates. Monocytes (MN) of patients with atherosclerosis and cardiac ischemia, undergoing stenting and conventional anti-atherosclerotic therapy, were harvested prior and after stenting. MN were isolated and cultured, with the transformation into MPh in direct contact with biopolymer culture substrates with different monomer composition and nano-reliefs, and transformed into MPh, in comparison with the same process on standard culture plastic. Sub-populations of cells with characteristic morphology in each phenotypic class were described, and their quantitative ratios for each sample of polymers were counted as an intermediate result in the development of “smart” material for cardiovascular devices. The results obtained allow us to assume, that the processes of MPh differentiation and polarization in vitro depend not only on the features of the micro/nano relief of biopolymer substrates, but also on the initial state of MN in vivo and general response of patients. © 2018 The Authors

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Держатели документа:
Siberian Federal University, 79, Svobodny av., Krasnoyarsk, 660041, Russian Federation
Institute of Biophysics, Siberian Branch of the Russian Academy of Sciences, 50/50 Akademgorodok, Krasnoyarsk, 660036, Russian Federation
L.V. Kirensky Institute of Physics, Siberian Branch of the Russian Academy of Sciences, 50/38 Akademgorodok, Krasnoyarsk, 660036, Russian Federation
Federal Research Center Krasnoyarsk Scientific Center of the Siberian Branch of the Russian Academy of Sciences, 50 Akademgorodok, Krasnoyarsk, 660036, Russian Federation
Federal Center for Cardiovascular Surgery, 45 Karaulnaya, Krasnoyarsk, 660020, Russian Federation

Доп.точки доступа:
Menzyanova, N. G.; Pyatina, S. А.; Nikolaeva, E. D.; Shabanov, A. V.; Nemtsev, I. V.; Stolyarov, D. P.; Dryganov, D. B.; Sakhnov, E. V.; Shishatskaya, E. I.

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