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Каримов И.Ф.
Samkova E.B., Karimov I.F.
ASSESMENT OF LIGHTING SOURCES BIOSAFETY BY REPORTER LUMINESCENT BACTERIA
Abstract. In recent years a number of studies purpose was to develop a biosensor methods for the evaluation of mutagenic and carcinogenic effects of environmental factors on the cell, in particular biosensor test systems whole cells that have in their composition two main genetic elements: a promoter that responds to exposure to a toxicant, and gene-reporter transcribed from the promoter. It may be noted that reporter microorganisms very suitable object for the study of DNA-damaging effects, due to ease of cultivation, inexhaustible resources, to ease of changing the irradiation conditions, and finally the ability to transcriptional response that is easy to visualize the level of luminescence of the bacterial strain. In this work reporter luminescent strains that are based on E. coli K12 MG1655: E. coli K12 MG1655 recA::lux, E. coli K12 MG1655 colD::lux and E. coli K12 MG1655 ibpA::lux were used to assess the biosafety sources artificial light, with the first two luminescence respond to DNA damage, and the latter — on the damage of heat shock proteins. For creating optimal conditions for the work of strains was established method of estimating the light, and a comparison of the physical characteristics of light sources, including incandescent lamps, compact fluorescent lamps and LED lamps. The results allowed to state that all types of lamps are able to provide the DNA damaging effects in different degrees, but more important is the brand, rather than the type of lamp; wherein a correlation damage from ultraviolet radiation intensity emitted by all types of light sources. There was also a difference in the response of the features of the strains E. coli recA::lux and E. coli colD::lux.
Key words: reporter strains, compact fluorescent lamp, incandescent, LED lamp, ultraviolet, DNA damage.
Drozdova E.A., Karimov I.F.
RUMEN FLUID NORMAL COMPONENTS EFFECTS ON LUMINESCENCE LEVEL OF BIOSENSOR
The effect of real (native) and rumen fluid mixture model on the emission level of luminescent strain E.coli was considered. Revealed similar toxic effects studied biological fluids, developing in the first minutes of contact. Analyzed the impact of individual components on rumen fluid bacteria glow, as well as the contribution of individual components of the trophic substrate.
Key words: biosensor, biotoxicity, rumen fluid, a recombinant strain of E. coli K12 TG1, substrate.
Karimov I.F., Manukhov I.V., Ivanov Y.B., Deryabin D.G.
BACILLUS SUBTILIS VKPM V-10548 STRAIN AND IT APPLICATION FOR SERUM BLOOD BACTIRICIDAL ACTIVITY RESEARCH
The recombinant Bacillus subtilis strain carrying plasmid with gram-negative sea microorganism Photobacterium leiognathi luxAB genes, before each introduced gram-positive bacteria ribosome-binding site is designed. This strain characterized by high gene expression level of bacterial luciferase and was deposited in the Russian National Collection of Industrial Microorganisms under № V-10548. In researching of B.subtilis VKPM V-10548 reaction in contact with different component serum blood probes the correspondence between bioluminescence suppression and bactericidal effect level mainly determined by beta-lysine activity is shown.
Key words: bioluminescence, lux-genes, Bacillus subtilis, bactericidal effect, beta-lysine
Karimov I.F., Ivanov Yu.B., Deryabin D.G.
INFLUENCE OF THROMBOCYTE CATION ALBUMEN ON BIOLUMINESCENCE AND VIABILITY OF RECOMBINANT STRAIN ESCHERICHIA COLI WITH CLONED LUX-OPERON PHOTOBACTERIUM LEIOGNATHI
High sensibility of bioluminescence response of recombinant strain E.coli K 12 TG1 with cloned luxCDABE with gene P.leiognathi 54D10 to the influence of thrombocyte cation albumen (TCA) is established in this article. Determined for TCA quantities IC50, corresponding to 50-percent inhibition of luminescence intensity from starting level form in the same system 3,3-7,1 mkg/ml. It is shown that concentration dependences of bioluminescence suppression at the presence of TCA are inferior a lot to the development true bactericidal effect regarding to used bacterial cell-targets. As one of the possible reasons of such phenomenon it was named direct abscopal influence of TCA on enzymatic system of luminescence generation.
Deryabin D.G., Karimov I.F.
PECULIARITIES RECOMBINANT LUMINESCENT BACTERIUM REACTION WITH DIFFERENT LUX-OPERONS IN PHAGOCYTAL SYSTEM
Dependence of bioluminescent response of different recombinant strains Escherichia coli in phagocytal system from the nature of cloned lux-operons in them is determined in this article. It is shown that constructions’ using on the base of genetic material of sea luminescent bacterium Photobacterium leiognathi in interaction with neutrophils of peripheral human blood allows reaching direct proportionality between light intensity and evidence of killing effect developed in these conditions.
Deryabin D.G., Grudinin D.A., Karimov I.F.
BASING OF OPTIMAL REGIME OF RECOMBINANT LUMINESCENT BACTERIA’S OPSONIZATION
Optimal procedure of recombinant strain Escherichia coli K12 having luxCDABE gene P.leiognathi with using of normal human immunoglobulin in contrast to other potential opsonines (intact and thermoinactivated blood serum, complement) not giving bactericidal effect and minimal changed level of its initial bioluminescence concerning this microorganism is based in this article. It is shown that recombinant strains E.coli opsonizated with given way cause dose-dependent stimulation of luminol-dependent chemiluminescence of neutrophilic phagocytes but at the same contact themselves demonstrate progressive decrease of bioluminescence level the nature of which is determined admittedly with absorption and intracellular killing of bacterial cells.
Deryabin D.G., Polyakov E.G., Karimov I.F.
PECULIARITIES OF USE OF BIOLUMINESCENT TEST - SYSTEMS BY RESEARCH OF ABIOTIC AND BIOLOGICAL LIQUIDS.
Comparative analysis on the basis of natural and recombinant luminous bacteria is conducted, and also luciferase systems detailed from them, by the research of abiotic spheres and biological liquids. Peculiarities of reaction of bioluminescence on influence of serum of blood, urine, bile, backbrain and other liquids are demonstrated, and it is the basis for development of new trend of usage of bioluminescent analysis in biology and medicine.
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Editor-in-chief |
Sergey Aleksandrovich
MIROSHNIKOV |
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