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1.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Sushchik N.N., Gladyshev M.I., Kalachova G.S., Guseynova V.E.
Заглавие : Rapid assay of fatty acid composition using a portable high-performance liquid chromatograph for monitoring aquatic ecosystems
Место публикации : Journal of Chromatography A. - 1995. - Vol. 695, Is. 2. - С. 223-228. - ISSN 00219673 (ISSN) , DOI 10.1016/0021-9673(94)01090-2
Ключевые слова (''Своб.индексиров.''): fatty acid--alga--article--culture medium--ecology--high performance liquid chromatography--instrument--methodology--priority journal--ultraviolet spectrophotometry--water analysis
Аннотация: The chromatographic conditions presented allowed the separation of the nitrophenacyl derivatives of standards of eleven free fatty acids (FFA) using a portable high-performance chromatograph, suitable for use aboard a research vessel. A statistically significant linear correlation between UV absorbance and amount of the analytes injected was obtained. The method was tested on FFA from algae cultural media. The method can be used for the ecological monitoring of natural waters.
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2.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Kratasyuk V.A., Egorova O.I., Esimbekova E.N., Kudryashova N.S., Orlova N.Y., L'vova L.S.
Заглавие : A biological luciferase test for the bioluminescent assay of wheat grain infection with Fusarium
Колич.характеристики :3 с
Место публикации : Appl. Biochem. Microbiol.: MAIK NAUKA/INTERPERIODICA, 1998. - Vol. 34, Is. 6. - P622-624. - ISSN 0003-6838
Примечания : Cited References: 7
Аннотация: The extent of inhibition of the bioluminescence reaction by wheat grain extracts was studied as a function of the scabby kernel content in wheat. The NADH : flavine mononucleotide oxidoreductase-luciferase bienzyme bioluminescence system was found to be the most sensitive to mycotoxins produced by fungi of the genus Fusarium. A biological luciferase test was developed for monitoring wheat grain infection with Fusarium.
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3.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Kuznetsov A.M., Rodicheva E.K., Medvedeva S.E.
Заглавие : Analysis of river water by bioluminescent biotests
Место публикации : Luminescence. - 1999. - Vol. 14, Is. 5. - С. 263-265. - ISSN 15227235 (ISSN)
Ключевые слова (''Своб.индексиров.''): bioluminescent bioassay--pollution--water quality--fresh water--article--bioassay--escherichia coli--freeze drying--genetic procedures--luminescence--methodology--photobacterium--russian federation--sensitivity and specificity--water pollutant--biological assay--biosensing techniques--escherichia coli--freeze drying--fresh water--luminescence--photobacterium--russia--sensitivity and specificity--water pollutants, chemical
Аннотация: The bacterial bioluminescence has high sensitivity to the action of various inhibitors of biological activity. The lyophilized luminous bacteria Photobacterium phosphoreum (Microbiosensor B17 677F) and luminous strain Escherichia coli (Microbiosensor EC) from the Culture Collection IBSO were used to create bioluminescent biotests. They have been applied in ecological monitoring to determine the overall toxicity of the Yenisei and Angara Rivers and some water sources of Altai Territory. As a rule the heaviest pollution of water in studied rivers was registered near cities and settlements. The luminous bacteria biotests are simple and convenient in work, standardized and quantitative, have rapid response to actions of different substances and high sensitivity to environmental pollutants. It takes less than 30 min to do the biotest (the other biotests take 48-96 h). Copyright В© 1999 John Wiley & Sons, Ltd.
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4.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Kratasyuk V.A., Esimbekova E.N., Gladyshev M.I., Khromichek E.B., Kuznetsov A.M., Ivanova E.A.
Заглавие : The use of bioluminescent biotests for study of natural and laboratory aquatic ecosystems
Место публикации : Chemosphere. - 2001. - Vol. 42, Is. 8. - С. 909-915. - ISSN 00456535 (ISSN) , DOI 10.1016/S0045-6535(00)00177-6
Ключевые слова (''Своб.индексиров.''): alcohol dehydrogenase--bacterial luciferase--bioluminescence--blooming--pollution--trypsin--water toxicity--alcohol dehydrogenase--benzoquinone--luciferase--trypsin--aquatic ecosystem--bioluminescence--water quality--article--bacterium culture--bioluminescence--blue green alga--ecosystem--pond--seasonal variation--water pollution--water quality--benzoquinones--biological assay--cyanobacteria--ecosystem--environmental monitoring--eutrophication--fmn reductase--indicators and reagents--luminescent measurements--nadh, nadph oxidoreductases--water pollutants--russian federation--algae--bacteria (microorganisms)--chlorophyta--cyanobacteria--uncultured cyanobacterium
Аннотация: A set of bioluminescent tests was developed to monitor water quality in natural and laboratory ecosystems. It consisted of four bioluminescent systems: luminous bacteria, coupled enzyme system NADH:FMN-oxidoreductase-luciferase and triplet enzyme systems with alcohol dehydrogenase and trypsin. The set of biotests was applied for a small forest pond (Siberia, Russia), laboratory microecosystems polluted with benzoquinone and a batch culture of blue-green algae. Thereby effects of natural water compared to those of models of heavy pollution and "bloom" of blue-greens on the bioluminescent tests were revealed. The set of biotests was not affected by a natural seasonal variability of water quality in the unpolluted pond, but responded to the heavy pollution and the "bloom" of blue-greens. The set of biotests could be recommended as the alarm test to control the acute toxicity of natural water bodies. В© 2001 Elsevier Science Ltd.
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5.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Remmel N.N., Titova N.M., Kratasyuk V.A.
Заглавие : Oxidative stress monitoring in biological samples by bioluminescent method
Место публикации : Bulletin of Experimental Biology and Medicine. - 2003. - Vol. 136, Is. 2. - P209-211. - ISSN 00074888 (ISSN) , DOI 10.1023/A:1026347830283
Ключевые слова (''Своб.индексиров.''): bioluminescent analysis--lipid peroxidation--malonic dialdehyde--oxidative stress--malonaldehyde--animal tissue--article--bioluminescence--brain tissue--controlled study--fluorescence--freeze drying--kidney parenchyma--lipid peroxidation--liver--male--monitoring--nonhuman--oxidative stress--photobacterium phosphoreum--rat--serum--animals--bacteria--biological assay--lipid peroxidation--luminescent measurements--malondialdehyde--oxidative stress--photobacterium--rats--tissue extracts--animalia--negibacteria--photobacterium--photobacterium phosphoreum
Аннотация: The integral bioluminescent biotest with lyophilized fluorescent bacteria was used for monitoring of LPO processes in tissue extracts and serum of rats exposed to stress. A relationship between the content of MDA (LPO indicator) and fluorescence of bacteria was observed in all biological samples.
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6.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Sid'ko A.F.
Заглавие : Remote assay for chlorophyll photosynthetic potential of crops on the example of wheat
Место публикации : Biol. Bull: MAIK NAUKA/INTERPERIODICA, 2004. - Vol. 31, Is. 5. - С. 450-456. - 7. - ISSN 1062-3590, DOI 10.1023/B:BIBU.0000043769.80086.02
Примечания : Cited References: 20
Аннотация: An optical remote assay for biological production of crops in the field during the vegetation period is proposed. Our calculations demonstrate a good correlation between the S(t) value and crop yield (in the range from 0.85 to 0.90); the higher is S(t) for the vegetation period, the higher is the crop yield. S(t) value can be used to calculate the increase in dry and wet biomass for various crops during the whole vegetation period with an error of less than 10-12%. The absolute error of crop yield for 110 cultivars of wheat, oats, and barley during the experimental period was +/-3-3.5 quintal/ha with the mean crop yield of 14-38 quintal/ha.
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7.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Frank L.A., Petunin A.I., Vysotski E.S.
Заглавие : Conjugates of the Ca2+-regulated photoprotein obelin with immunoglobulins: Synthesis and use as labels in bioluminescent immunoassay
Колич.характеристики :5 с
Место публикации : Russ. J. Bioorg. Chem.: MAIK NAUKA/INTERPERIODICA, 2004. - Vol. 30, Is. 4. - С. 327-331. - ISSN 1068-1620, DOI 10.1023/B:RUBI.0000037257.80835.7a
Примечания : Cited References: 16
Предметные рубрики: ESCHERICHIA-COLI
PURIFICATION
AEQUORIN
PROTEIN
CDNA
Ключевые слова (''Своб.индексиров.''): bioluminescent immunoassay--obelin--thyroid stimulating hormone
Аннотация: An efficient procedure for obelin conjugation with immunoglobulins was developed. The possibility was shown of using the resulting conjugates instead of a radioisotope label for the immunoassay of thyroid stimulating hormone in sera; the conjugates provide a sensitivity of 0.01 muIU/ml. The results of bioluminescent immunoassay (sera of 34 patients) satisfactorily correlate with the results of radioisotope assay (R 0.99).
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8.

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.
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9.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Sid'ko A.F.
Заглавие : Remote assay for chlorophyll photosynthetic potential of crops on the example of wheat
Колич.характеристики :7 с
Место публикации : Biol. Bull: MAIK NAUKA/INTERPERIODICA, 2004. - Vol. 31, Is. 5. - P450-456. - ISSN 1062-3590, DOI 10.1023/B:BIBU.0000043769.80086.02
Примечания : Cited References: 20
Аннотация: An optical remote assay for biological production of crops in the field during the vegetation period is proposed. Our calculations demonstrate a good correlation between the S(t) value and crop yield (in the range from 0.85 to 0.90); the higher is S(t) for the vegetation period, the higher is the crop yield. S(t) value can be used to calculate the increase in dry and wet biomass for various crops during the whole vegetation period with an error of less than 10-12%. The absolute error of crop yield for 110 cultivars of wheat, oats, and barley during the experimental period was +/-3-3.5 quintal/ha with the mean crop yield of 14-38 quintal/ha.
WOS
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10.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Frank L.A., Petunin A.I., Vysotski E.S.
Заглавие : Conjugates of the Ca2+-regulated photoprotein obelin with immunoglobulins: Synthesis and use as labels in bioluminescent immunoassay
Колич.характеристики :5 с
Место публикации : Russ. J. Bioorg. Chem.: MAIK NAUKA/INTERPERIODICA, 2004. - Vol. 30, Is. 4. - P327-331. - ISSN 1068-1620, DOI 10.1023/B:RUBI.0000037257.80835.7a
Примечания : Cited References: 16
Предметные рубрики: ESCHERICHIA-COLI
PURIFICATION
AEQUORIN
PROTEIN
CDNA
Ключевые слова (''Своб.индексиров.''): bioluminescent immunoassay--obelin--thyroid stimulating hormone
Аннотация: An efficient procedure for obelin conjugation with immunoglobulins was developed. The possibility was shown of using the resulting conjugates instead of a radioisotope label for the immunoassay of thyroid stimulating hormone in sera; the conjugates provide a sensitivity of 0.01 muIU/ml. The results of bioluminescent immunoassay (sera of 34 patients) satisfactorily correlate with the results of radioisotope assay (R 0.99).
WOS
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11.

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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12.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Shishatskaya E.I., Khlusov I.A., Volova T.G.
Заглавие : A hybrid PHB-hydroxyapatite composite for biomedical application: Production, in vitro and in vivo investigation
Место публикации : Journal of Biomaterials Science, Polymer Edition. - 2006. - Vol. 17, Is. 5. - С. 481-498. - ISSN 09205063 (ISSN) , DOI 10.1163/156856206776986242
Ключевые слова (''Своб.индексиров.''): biocompatibility--hydroxyapatite (ha)--phb-hydroxyapatite composite--polyhydroxyalkanoate (pha)--polyhydroxybutyrate (p(3hb))--properties--biocompatibility--differential thermal analysis--electron microscopy--free energy--interfacial energy--physical properties--surface properties--x ray analysis--biomedical application--physicochemical properties--polyhydroxyalkanoate (pha)--polyhydroxybutyrate (phb)--hydroxyapatite--hydroxyapatite--poly(3 hydroxybutyric acid)--polymer--biomaterial--hydroxybutyric acid--adhesion--animal cell--animal tissue--article--biomedicine--bone marrow cell--cell differentiation--cell growth--chemical structure--composite material--controlled study--crystallization--decomposition--electron microscopy--in vitro study--in vivo study--melting point--mouse--nonhuman--ossification--osteoblast--physical chemistry--priority journal--rat--strength--structure analysis--surface property--synthesis--temperature measurement--thermal analysis--tissue engineering--wettability--animal--biomechanics--bioremediation--bone prosthesis--cattle--cell culture--chemistry--cytology--differential scanning calorimetry--drug effect--human--materials testing--prostheses and orthoses--scanning electron microscopy--standard--wistar rat--murinae--animals--biocompatible materials--biodegradation, environmental--biomechanics--bone substitutes--cattle--cells, cultured--differential thermal analysis--durapatite--humans--hydroxybutyrates--materials testing--microscopy, electron, scanning--osteoblasts--prostheses and implants--rats--rats, wistar--surface properties
Аннотация: Samples of a hybrid composite of polyhydroxybutyrate (PHB), a biodegradable polyester, and hydroxyapatite (HA), with different PHB/HA ratios, have been prepared using mechanical-physical method. Electron microscopy, X-ray structure analysis and differential thermal analysis have been used to investigate the structure and physicochemical properties of the composite, depending on the PHB/HA ratio. The properties of the surface of the HA-loaded composite are significantly different from those of the pure polymer. As the HA percentage in the composite increases, free interface energy, the cohesive force, i.e., the strength of the adhesive bond between the composite surface and the water phase, and surface wettability increase. The HA percentage of the composite does not influence its melting temperature, but affects the temperature for the onset of decomposition: as the HA content increases from 0 to 10% (w/w), Td decreases from 260В°C to 225В°C. The degree of crystallinity of PHB/HA increases from 77% to 89% with an increase in the HA fraction from 10% to 50%. Functional properties of the composites have been investigated in vitro and in vivo. The best parameters of growth and differentiation of murine marrow osteoblasts are registered on PHB/HA samples containing 10% and 20% HA. In ectopic bone formation assay it has been proven that the hybrid PHB/HA composites can function as scaffolds and that bone tissue develops on their surface and in pores. В© VSP 2006.
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13.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Rozhko T.V., Kudryasheva N.S., Kuznetsov A.M., Vydryakova G.A., Bondareva L.G., Bolsunovsky A.Y.
Заглавие : Effect of low-level alpha-radiation on bioluminescent assay systems of various complexity
Место публикации : Photochem. Photobiol. Sci.: ROYAL SOC CHEMISTRY, 2007. - Vol. 6, Is. 1. - С. 67-70. - 4. - ISSN 1474-905X, DOI 10.1039/b614162p
Примечания : Cited References: 52
Предметные рубрики: LOW-DOSE RADIATION
IRRADIATION
TOXICITY
QUINONES
HORMESIS
PHENOLS
Аннотация: This study addresses the effects of low-level alpha-radiation on bioluminescent assay systems of different levels of organization: in vivo and in vitro. Three bioluminescent assay systems are used: intact bacteria, lyophilized bacteria, and bioluminescent system of coupled enzyme reactions. Solutions of Am-241(NO3)(3) are used as a source of alpha-radiation. It has been shown that activation processes predominate in all the three bioluminescent assay systems subjected to short-term exposure (20-55 h) and inhibition processes in the systems subjected to longer-term exposure to radiation. It has been found that these effects are caused by the radiation component of Am-241(3+) impact. The intensity of the Am-241(3+) effect on the bioluminescent assay systems has been shown to depend on the Am-241(3+) concentration, level of organization and integrity of the bioluminescent assay system. The bioluminescent assay systems in vivo have been found to be highly sensitive to Am-241(3+) (up to 10(-17) M).
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14.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Frank L..., Markova S..., Remmel N..., Vysotski E..., Gitelson I...
Заглавие : Bioluminescent signal system: bioluminescence immunoassay of pathogenic organisms
Колич.характеристики :6 с
Место публикации : Luminescence: JOHN WILEY & SONS LTD, 2007. - Vol. 22, Is. 3. - С. 215-220. - ISSN 1522-7235, DOI 10.1002/bio.952
Примечания : Cited References: 14
Предметные рубрики: AEQUORIN
AGENTS
OBELIN
ASSAYS
LABEL
Ключевые слова (''Своб.индексиров.''): obelin--bioluminescence immunoassay--infective agents
Аннотация: The Ca2+-regulated photoprotein obelin has been examined as a label for bioluminescence immunoassay of infective agents. The hepatitis B virus (HbsAg) and the bacteria Escherichia coli and Shigella sonnei lipopolysaccharide (LPS) were chosen as model antigens. Chemically synthesized obelin-corresponding antibody conjugates were used in a solid-phase microplate immunoassay. The sensitivities achieved by the assay were 0.25 ng/mL for S. sonnei LPS and 0.375 ng/mL for HbsAg. A novel, filter-based immunoassay to determine bacterial admixtures in the environment was proposed. The NanoCeram filters were effectively applied to 'trap' and pre-concentrate pathogens from samples under study for the purposes of further detection and measurement of the absorbed material by bioluminescence immunoassay. Copyright (C) 2007 John Wiley & Sons, Ltd.
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15.

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.
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16.

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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17.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Shishatskaya E.I.
Заглавие : Biomedical investigations of biodegradable PHAs
Место публикации : Macromolecular Symposia. - 2008. - Vol. 269, Is. 1. - С. 65-81. - ISSN 10221360 (ISSN) , DOI 10.1002/masy.200850909
Ключевые слова (''Своб.индексиров.''): biodegradable polymers--biomedical investigations--pha--abs resins--biodegradable polymers--biopolymers--biotechnology--bone--cell culture--endothelial cells--fibers--functional polymers--osteoblasts--polymers--surgery--abdominal surgeries--biocompatible--biodegradable--biomedical--biomedical investigations--bone defects--ectopic bones--hepatocytes--in vitro--microparticles--oral surgeries--osteogenesis--pha--polyhydroxybutyrate--polymer devices--ralstonia--russian academy of sciences--two types--polymer films
Аннотация: This work is a review of the results of biomedical studies of polymer devices (films, fibers, microparticles, 30 implants) made from resorbable PHAs synthesized by the bacterium Wautersia (Ralstonia) eutropha 65786, using the technology developed at the Institute of Biophysics of the Siberian Branch of the Russian Academy of Sciences. Two types of PHAs - polyhydroxybutyrate (PHB) and a hydroxybutyrate/hydroxyvalerate copolymer (PHB/PHV) - have been proven to be biocompatible in vitro in cultures of fibroblasts, endothelial cells, hepatocytes, and osteoblasts, and in short- and long-duration experiments on animals. Polymer films and membranes have been found to be usable as scaffolds for functioning cells and monofilament suture fibers - for stitching muscular-fascial wounds and in abdominal surgery. Ectopic bone formation assay and experiments with the model of segmental osteotomy showed that 30 PHB and PHB/HA implants can be used for reparative osteogenesis. The paper reports beneficial results of using polymers to repair bone defects in oral surgery. Copyright В© 2008 WILEY-VCH Verlag GmbH & Co. KGaA.
Scopus
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18.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Borisova V.V., Frank L.A., Korneeva S.I., Vysotski E.S.
Заглавие : Metridia luciferase and Renilla Ca2+-dependent coelenterazine-binding protein as reporters for in vitro assay
Колич.характеристики :1 с
Место публикации : Luminescence: JOHN WILEY & SONS LTD, 2008. - Vol. 23, Is. 2. - С. 60-60. - ISSN 1522-7235
Примечания : Cited References: 0
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19.

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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20.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Frank L.A.
Заглавие : Ca2+-Regulated Photoproteins: Effective Immunoassay Reporters
Колич.характеристики :14 с
Коллективы :
Место публикации : Sensors: MDPI AG, 2010. - Vol. 10, Is. 12. - С. 11287-11300. - ISSN 1424-8220, DOI 10.3390/s101211287
Примечания : Cited References: 70. - This work was supported by the grant No. 76 of the Russian Academy of Sciences, Siberian Branch.
Предметные рубрики: POLYMERASE-CHAIN-REACTION
CYTOKINE MESSENGER-RNA
BIOLUMINESCENT IMMUNOASSAY
MYCOBACTERIUM-TUBERCULOSIS
BIOTINYLATED AEQUORIN
RECOMBINANT AEQUORIN
ANGSTROM RESOLUTION
FUSION PROTEIN
PCR ASSAY
OBELIN
Ключевые слова (''Своб.индексиров.''): bioluminescence--ca2+-regulated photoprotein--immunoassay--pcr-elisa--multiplex assay--re-engineered photoproteins
Аннотация: Ca2+-regulated photoproteins of luminous marine coelenterates are of interest and a challenge for researchers as a unique bioluminescent system and as a promising analytical instrument for both in vivo and in vitro applications. The proteins are comprehensively studied as to biochemical properties, tertiary structures, bioluminescence mechanism, etc. This knowledge, along with available recombinant proteins serves the basis for development of unique bioluminescent detection systems that are "self-contained", triggerable, fast, highly sensitive, and non-hazardous. In the paper, we focus on the use of photoproteins as reporters in binding assays based on immunological recognition element-bioluminescent immunoassay and hybridization immunoassay, their advantages and prospects.
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