Addition of medicinal plant Ungernia victo ris extract to bacterial nutrient medium rises significantly yield of cell biomass and target protein of strains ITG1 (producer of recombinant NH 2terminal fragment of βgalactosidase) and ITG71Δ (producer of recombinant chimerical polypeptide with immunological properties of HIV protein Env1). When relatively poor LB medium is used, low concentrations of extract (0.25, 0.5 and 1%) do not ensure effect of stimulation. At the same time the presence of extract in higher concentrations (2.0, 5.0 and 10%) acts to rise further increasing of biomass level and total content of target protein on unit of medium volume for both strains. So the results of experiments carried with use of LB medium demonstrate principle ability of U. victoris extract for raising final yield of target product at the expense of two mechanisms — increase of biomass yield and growth of percentage of recombinant polypeptide in total cell protein. Unlike the results obtained in LB medium, extract of 2.0, 5.0 and 10% concentrations in enriched medium that is in use usually for industrial cultivation of strainsproducers demonstrates even stable tendency to inhibition of final yield of biomass and target polypeptide aside from stimulate. For all that addition of U. victoris extract to enriched medium in more low concentrations (0.25–1%) results as a rule in significant yield increasing of target proteins for both strains. Data obtained correspond to our data received earlier for plasmidless strains and prove general regularities according to that ones addition of medicinal plant extract in high concentrations to poor medium and in low concentrations to enriched one facultates significant rise of biomass and target protein yields per unit volume of cultivation medium.

Optimization of culture media is considered to be one of the ways to increase biomass output of bacterial cells as well as a base product at cultivation of all types and purposes. In connection with the development of the biotechnology industrya, search of the new components of nutrient solution and their optimum ratio with the base composition of the medium and corresponding properties of the cultivated object is becoming increasingly important. Addition of Ungernia victoris extract to bacterial nutrient medium optimizes its composition and enables to increase production of E. coli biomass. Statistical reliable exceeding of biomass production of M17 and HB101 strains in LB medium occurs only at higher concentrations of extract (5–10%) while biomass production of JM109 strain increases at all tested concentrations (0,5–10%). The highest biomass yield of all three strains corresponds to extract concentration of 10%. The influence of U. victoris extract on growth of bacterial strains in enriched medium differs from its influence on the same strains in LB medium. Biomass increase happens at low concentrations of extract (0.25–1%)and it is not such significant as its increase in LB medium. Starting with extract concentration from 2% and to 10%, the tendency towards biomass decrease for M17 and HB101 strains and lower biomass increase for JM109 strain as compared with 0.25–1% concentrations takes place. Thus, it is possible to get improvement of bacterial biomass production by means of plant extract using both poor and enriched medium. Optimal quantity of the added extract fluctuates depending on medium composition and peculiarities of object of growth.

Components of three investigated plant extracts from the biomass of cultivated cells of Ungernia victoris, Rhodiola rosea, and Polyscias filicifolia interact with porin proteins OmpC and OmpF and reduce their receptor activity towards OmpC- and OmpF-dependent bacteriophages. The influx of these components into cells optimizes the synthesis of a lipopolysaccharide (LPS) complex and contributes to the enhancement of receptor activity of not only LPS but also OmpA and LamB proteins closely associated with the latter.

Sensitive cells of Escherichia coli AB 259 Hfr 3000 infected with RNA-containing phage MS2 produce phage particles and simultaneously continue to divide, thereby segregating sensitive cells capable of sustaining new cycles of infection. Multiplication of phage in sensitive cells gives rise to phage-resistant forms in the progeny of these cells. It is shown that this phenomenon is due not to selection of pre-existing phage-resistant mutants, but is instead the result of interaction between the phage and the cell. Unlike ordinary spontaneous or chemically induced E. coli mutants, MS2-induced phage-resistant cells are genetically unstable forms. In the course of reproduction they segregate new MS2-resistant forms with more highly expressed variations in the region encoded by the sex factors. Cells of the two final forms of MS2-induced mutants also produce a new type of phage. This new type constitutes DNA-containing forms which, however, are neutralized by anti-MS2-serum. The segregation of these forms serves to confirm that the genetic substance of the RNA-containing bacteriophage is capable of being expressed as a component of the DNA-containing structure.

Furosemid and Verapamil influence the production of plasmid transformants in E. coli in accordance with the actual properties of the calcium channel regulators in eukaryotes. That is, Furosemid stimulates, while Verapamil inhibits, entry of transforming DNA into a competent cell. The PHB/Ca2+ polyP complex not only serves as a channel for penetration of transforming DNA, but also functions as one of the targets for blocking transformation of a U. victoris extract.

A fundamental possibility of antitumor activity testing in the bacterial system represented by Escherichia coli of the wild type and its MS2-induced mutant has been shown. The initial bacterial strain is an indicator of toxic properties of tested substances and the mutant strain is a specific test culture modeling a tumor cell. The comparison of the data described for eukaryotes with the data obtained using the proposed bacterial test system confirms an adequate response of both strains to substances with proved antitumor properties. The data obtained are considered as very promising for the further improvement of this test system towards its application for primary screening of antitumor substances.

A principle possibility of antitumour activity test in bacterial system represented by Escherichia coil of the wild type and its MS2-induced mutant has been shown. The initial bacterial strain is an indicator of toxic properties of the tested substances and the mutant one is a specific test-culture modelling a tumour cell. The comparison of the data described for eucaryotes with the data obtained using the proposed bacterial test system confirms an adequate response of both strains to the substances with the proved antitumour properties. The data are considered as very promising for the further improvement of this test system towards its application for primary screening of antitumour substances.

Ability of cell biomass extracts of some medicinal plants to inhibit spontaneous segregation of new forms in E. coli MS2-induced unstable mutants system has been shown. The effect described is analyzed from the position of stabilizing activity of extracts concerning bacterial envelope membranes. The possible use of this bacterial system to study stabilizing effect as a separate stage of the known antimutagen protective action is discussed.

In the system of cation-induced E. coli transformation by the plasmid pBR322 the effects of the extract derived from the biomass of cultured cells of U. victoris on the correlation between yield of transformants, viability of CaCl2 - treated cells and plasmid DNA conformation alterations has been investigated. The data obtained have been compared with effects of some salts of one- and divalent metals on the same parameters. The presence of different mechanisms of the variations of cell population viability and yield of transformants depending on utilization of salts or plant extracts has been shown.

Conformation analysis of mutated DNA-bacteriophages (PLys-23, P23-2, P47- the numbers have been assigned by T. Pererva) induced by MS2 virus incorporated in E. coli AB 259 Hfr 3000 has been done. Surface enhanced infrared absorption (SEIRA) spectroscopy and principal component analysis has been applied for solving this problem. The nucleic acids isolated from the mutated phages had a form of double stranded DNA with different modifications. The nucleic acid from phage P47 was undergone the structural rearrangement in the most degree. The shape and position of the fine structure of the Phosphate asymmetrical band at 1071cm(-1) as well as the stretching OH vibration at 3370-3390 cm(-1) has indicated to the appearance of additional OH-groups. The Z-form feature has been found in the base vibration region (1694 cm(-1)) and the sugar region (932 cm(-1)). A supposition about modification of structure of DNA by Z-fragments for P47 phage has been proposed. The P23-2 and PLys-23 phages have showed the numerous minor structural changes also. On the basis of SEIRA spectra we have determined the characteristic parameters of the marker bands of nucleic acid used for construction of principal components. Contribution of different spectral parameters of nucleic acids to principal components has been estimated.

Antimutagenic system is represented as the system combining all enzymatic and metabolic processes of the organism at all levels. The problem of supporting of autoantimutagenic system with antimutagenic preparations is discussed. A number of possible mechanisms of antimutagenic action of different substances are considered.

In Escherichia coli--bacteriophage lambda system protective properties of the extracts derived from the biomass of cultured Panax ginseng, Polyscias filicifolia, Rhodiola rosea, Ungernia victoris cells, and those from intact Rhodiola roots have been studied. Escherichia coli--bacteriophage lambda system responsiveness was found to vary with the test-object state, namely: the deleted bacteriophage form (lambda-4) as well as previously mutagenized bacteriophage were more sensitive to the mutagenic and antimutagenic influence versus the native bacteriophage lambda +. The contribution of extracts in the induction and realization of the lethal injuries in phages caused by nitrite acid in extracellular phage (conditions in vitro) was estimated thus enabling to discriminate between the protective and antimutagenic extract activities. Protective extract effect in the given test-system appeared to be higher their antimutagenic action. With the most responsive bacteriophage variant the extracts from the biomass of cultured Rh. rosea and P. filicifolia cells showed high protective and somewhat lower antimutagenic activities. With other phages significant antimutagenic potential of extracts was demonstrated, which by their protective effect could be arranged in a raw as follows U. victoris > P. ginseng > P. filicifolia. The primary screening for the antimutagenic effect of preparations in the prokaryotic systems could be reduced to the investigation of their effects on the object inactivation exposed to the mutagen in vitro.

Using bacteriophages MS2, P17 and λ-vir infecting Escherichia coli AB 259 (Hfr 3000), E. coli Ml7 and E. coli C600 cells appropriately the author demonstrates that both RNA-and ONA-containing phages induce the development of phage-resistant forms appearing with high frequency in the progeny of infected cells.

The DNA-containing phages neutralizing with anti-MS2 serum have been detected in MS2-inducco bacterial, cultures, in the cells containing the recombinant plasmid as well as in preparations of transducing lambda and PI phages.The identity of all these new developed bacteriophages permits to suppose the common mechanism of their origin.

Some genotype and phenotype properties of MS2-induced being granular and lysing Escherichia coli mutants are described — form of colonies, cells form and size, susceptibility to lytic agents, carbohydrates utilization, antigenic properties, Gram staining, patterns of growth and division. The numerous aberrations from wild type bacteria found in these mutants are supposed to be due not to responsible loci mutations but to accompany structural rearrangements in cell membrane components.