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Deryabin D.G.,Orenburg State University | Karimov I.F.,Orenburg State University | Manukhov I.V.,State Institute of Genetics and Selection of Industrial Microorganisms | Tolmacheva N.A.,Orenburg State University | Balabanov V.P.,State Institute of Genetics and Selection of Industrial Microorganisms
Bulletin of Experimental Biology and Medicine | Year: 2012

Luminescence intensity of recombinant Escherichia coli and Bacillus subtilis strains with cloned luxCD(AB)E genes of the natural luminescent microorganism Photobacterium leiognathi was studied under the influence of 30 individual samples of human blood serum of different component composition. A relationship was found between the level of residual bioluminescence and degree of the bactericidal effect. Moreover, the inhibition of E. coli lux + luminescence was shown to be related to activity of the complement-lysozyme system. The reaction of B. subtilis lux + primarily depended on the presence of β-lysin in the blood serum. These data provide an experimental substantiation of a new method of differential analysis of humoral factors of nonspecific innate immunity with recombinant luminescent bacteria. © 2012 Springer Science+Business Media New York.


Zavilgelsky G.B.,State Institute of Genetics and Selection of Industrial Microorganisms | Kotova V.Y.,State Institute of Genetics and Selection of Industrial Microorganisms | Manukhov I.V.,State Institute of Genetics and Selection of Industrial Microorganisms
Nanotechnologies in Russia | Year: 2011

To investigate the mechanism of the bactericidal effect of TiO2 nanoparticles, we used induced lux biosensors on the basis of the Escherichia coli MG1655 strain containing pColD-lux, pKatG-lux, and pSoxS-lux plasmids. It was shown that the UV-light (λ = 360-390 nm) treatment of a suspension of biosensor cells and TiO2 nanoparticles induces H2O2 formation in bacteria, which induces the expression of the luxCDABE reporter genes, leading to bioluminescence intensification. It is proposed to use the specifically induced lux biosensors for the detection of photoactive dioxide metal nanoparticles. © 2011 Pleiades Publishing, Ltd.


Zavilgelsky G.B.,State Institute of Genetics and Selection of Industrial Microorganisms | Kotova V.Y.,State Institute of Genetics and Selection of Industrial Microorganisms | Rastorguev S.M.,State Institute of Genetics and Selection of Industrial Microorganisms
Russian Journal of Genetics | Year: 2011

The ArdA and Ocr antirestriction proteins, whose genes are in transmissible plasmids (ardA) and bacteriophage genomes (0. 3 (ocr)), specifically inhibit type I restriction-modification enzymes. The Ocr protein (T7 bacteriophage) was shown to inhibit both restriction (endonuclease) and modification (methylase) activities of the EcoKI enzyme in a broad range of intracellular concentrations (starting from 10-20 molecules per cell). In contrast to Ocr, the ArdA protein (ColIb-P9 transmissible plasmid) inhibited both of the EcoKI activities only at high intracellular concentrations (30000-40000 molecules per cell). When the ArdA concentration was several fold lower, only endonuclease activity of EcoKI was inhibited. It was assumed that a poorer ArdA ability to inhibit EcoKI modification activity is related to the substantial difference in life cycle between transmissible plasmids (symbiosis with the bacterial cell) and bacteriophages (infection and lysis of bacteria). The Ocr and ArdA mutants that inhibited exclusively endonuclease activity of EcoKI were obtained. Antirestriction proteins incapable of homodimerization were assumed to inhibit only endonuclease activity of type I restriction-modification enzymes. © 2011 Pleiades Publishing, Ltd.

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