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Applied Biochemistry and Microbiology (v.46, #9)

Antimicrobial peptides and their use in medicine by V. N. Lazarev; V. M. Govorun (pp. 803-814).

The review presents the current classification of antimicrobial peptides (AMP), which are the main component of innate immunity. The mechanism of their action and the molecular basis of the formation of resistance towards these peptides are described. Data on the use of AMP for the treatment of various infectious diseases, as well as the state of the art in genetic therapy using AMP, are given.

Keywords: antibiotic resistance; antimicrobial peptides; infectious diseases

Progress in the development of pandemic influenza vaccines and their production technologies by O. I. Kiselev (pp. 815-830).

This article analyzes the current situation in the field of construction and production of pandemic influenza vaccines. The main task of protecting the population against influenza pandemics requires state-of-the-art approaches to the construction of influenza vaccines to be based on reassortment and genetic engineering techniques, including the analysis of primary structures of influenza viral genes, synthesis and cloning of the main viral genes, reverse genetics techniques, and banks of plasmids bearing basic viral genes. Reassortant technologies are now giving way to new approaches for objective reasons. The state-of-the-art technologies provide safety not only at the laboratories where vaccine viruses are constructed but also make the production process wholly safe. We are using the following approaches to the development of industrial production: use of nanoparticles and nanoemulsions as functional adjuvants, construction of totally-safe strains for live attenuated influenza vaccines with deletions of molecular determinants of pathogenicity, application of protein and chemical chaperones to provide self-assembly of haemagglutinin molecules of the H1N1v-2009 virus, and impregnation of whole-virion preparations with nanoparticles to enhance antigenicity.

Keywords: adjuvants; antigens; vaccines; viral proteins; virus-like particles; live attenuated vaccines; replication-deficient vaccines

Effects of signal peptide primary structure on efficiency of recombinant Staphylococcus aureus pro-enterotoxin A transmembrane translocation in E. coli cells by V. A. Manuvera; N. N. Mordkovich; V. P. Veiko; V. G. Debabov (pp. 831-837).

Enterotoxin A containing various leader sequences have been obtained by site-driven mutagenesis. Some of them were capable of providing the translocation of recombinant SEA to E. coli periplasmic space. Structure of C-region of the signal peptide is essential for intracellular protein location. Substitution of a more common to E. coli proproteins Ala-Ser-Ala for the authentic sequence of Val-Asn-Gly is the most important in recognition by signal peptidase type I.

Keywords: E. coli ; enterotoxin; signal peptide; Staphylococcus aureus

Recombinant Cry9A is able to form crystals in sporulating cells of Bacillus thuringiensis by N. E. Kirillova; E. I. Levitin; Ya. A. Voytsekhovskaya; T. G. Yudina; I. A. Zalunin; G. G. Chestukhina (pp. 838-844).

An expression system for an effective production of recombinant protein Cry9A in bacillary cell has been suggested in the study. The proteins’ immunological properties, ability to proteolysis, and biological activity were identical to natural protein. The ability of recombinant Cry9A to form crystal bodies in sporulating cells of Bacillus thuringiensis has been shown. Thus, the first evidences of the fact that Cry-proteins which in natural strains form the crystal bodies together with other endotoxins are able to independently form the crystals has been received. The introduced system including vector replicative carriers, expression cassettes, and a protocol of obtaining and cultivation of strain-producer allows simple manipulations with the gene of delta-endotoxin of Cry9A in gene-engineering experiments.

Keywords: bioinsecticides; entomocidal crystals formation; recombinant protein; δ-endotoxins; Bacillus thuringiensis ; Cry9A

Peculiarities of luminescent response of Bacillus subtilis recombinant strain bearing cloned Vibro harveyi lux AB genes to the action of thermostable blood serum compounds by I. F. Karimov; I. V. Manukhov; V. Yu. Kotova; D. O. Omel’chenko; D. G. Deryabin (pp. 845-848).

A strain of Bacillus subtilis previously used as the test-organism in bacteriological and nephelometry methods for detection of thrombocytes cation protein (TCP) has been transformed by a plasmid pLFlux containing cloned luxAB genes of a sea luminescent bacterium Vibro harveyi. The designed luminescent biosensor B. subtilis of The All-Russia Collection of Industrial Microorganisms (Moscow) B-10191 demonstrated specific response to the thermostable fraction of guinea pig blood serum. Sensitivity to chromatographically purified TCP as part of the system has been analyzed and the correlation between luminescence inhibition and direct bacterial effect on the target cells has been demonstrated. The obtained results are considered as the first stage of the design of the bioluminescent technology for TCP detection in biological liquids with complicate composition of the components.

Keywords: bioluminescence; blood serum; thrombocytes cation protein; Bacillus subtilis ; Vibro harveyi

New DNA methyltransferase M.AjnI from the bacterium Acinetobacter johnsonii R2 produces the 5′-m5CCWGG-3′ sequence by V. S. Dedkov (pp. 849-853).

Optimum conditions for the activity of the new DNA methylase in cell lysate were determined. Methylation of DNAs of bacteriophages λ and T7 and plasmid pBR322 (dcm+) in the 5′-Cm5CWGG-3′ region blocked M.AjnI activity. The specificity of M.AjnI was determined using λ DNA methylated by this enzyme as well as computer modeling and data on the sensitivity of restriction endonucleases Mval, HinfI, and BstMAI to methylation.

Keywords: analysis of specificity; computer modeling; DNA-methyltransferase

Isolation and characterization of antibiotics produced by Streptomyces chrysomallus R-21 and S. globisporus L-242 strains by Yu. D. Shenin; I. I. Novikova; P. V. Suika Uarkaja (pp. 854-864).

Chrysomal and globerin complexes containing antibiotics from polyether and heptaene aromatic macrolide groups have been isolated from the mycelium of Streptomyces chrysomallus R-21 and Streptomyces globisporus L-242 strains, respectively. Physicochemical characteristics of these complexes have been investigated, including UV and NMR spectra, chromatographic parameters, etc. The fungicidal and antiviral activity of the studied complexes has been demonstrated on tomato plants. The obtained data allow us to consider the above-mentioned antibiotics to be original preparations.

Keywords: antibiotics; isolation; identification; model experiment; polyene; polyether; streptomycete; fungicidal activity

Development of technology for purification of plasmid DNAs for pharmaceutical purposes by E. A. Volosnikova; N. I. Akulova; Ya. S. Gogina; G. M. Levagina; V. K. Mikhailova; L. R. Lebedev; V. F. Podgornyi; Yu. V. Telegina (pp. 865-869).

Optimization of the stage of fractioning of nucleotide material has been performed, and a technology for purification and obtaining of plasmid DNAs for pharmaceutical purposes has been developed. Plasmid DNAs can be used for construction of DNA vaccines to urgent viral infections. Conditions for extraction and preliminary purification of DNA by salt and alcohol precipitations were determined. An effective gel-filtration procedure of separation of plasmid DNA from contaminants was suggested.

Keywords: plasmid DNAs; purification technology

Study of the sorption of glycopeptide antibiotics on an ultradisperse carbon sorbent by A. V. Timofeeva; V. N. Buravtsev; O. A. Galatenko; I. V. Tolstykh; L. P. Terekhova; A. V. Nikolaev; L. A. Baratova; G. S. Katrukha (pp. 870-878).

The sorption capacity of a novel ultradisperse carbon sorbent (UDCS) towards antibiotics of the glycopeptide group, namely, eremomycin, vancomycin, ristomycin A, and teicoplanin A2, has been studied. The conditions for maximum sorption of the antibiotics from solutions have been determined, and a mathematical model of the sorption of the antibiotics of the above named group has been put forward.

Keywords: glycopeptide antibiotics; quantitative analysis; Milikhrom A-02; RP-HPLC; ultradisperse carbon sorbent

Application of triplex PCR for identification of genetically modified organism in foods by E. S. Bulygina; M. V. Sukhacheva; B. K. Bumazhkin; B. B. Kuznetsov (pp. 879-883).

A multiplex method for detection of genetically modified organism (GMO) in various foods has been developed based on PCR-identification of cauliflower mosaic virus (CMV) 35S-promoter. It allows avoiding false positive signals due to contamination of plant raw material with CMV.

Keywords: foodstuff; GMO; GMO identification; PCR

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Applied Biochemistry and Microbiology (v.46, #9)

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Applied Biochemistry and Microbiology (v.46, #9)