Evaluation Of The Mutagenic Potential Of The Polyphenol Extract From Blueberries In The Ames Test

In this research, mutagenic properties of blueberry polyphenol extract were studied in gene mutation induction test (Ames test) on four strains of Salmonella typhimurium TA98, TA100, TA1535, TA1537. None of the strains of Salmonella typhimurium showed statistically reliable dose-dependent increase in number of revertant colonies in the presence of investigated drug in the studied dose range from 4,0 to 40,0 mg/ml relative to baseline of spontaneous mutations. The blueberry extract does not have any mutagenic activity in the researched dose range in relation to TA98, TA100, TA1535, TA1537 strains of Salmonella typhimurium.

Molybdoenzyme Participation In Plant Biochemical Processes

Molybdenum is a key microelement in plant vital functioning. The microelement can be absorbed by the plants only in the form of molybdate-anion. The Molybdenum deficiency affects negatively to the most important agricultural growing. As molybdenum takes part in such vital mechanisms as nitrogen and sulfur metabolism, plant hormone biosynthesis, and purine banding catabolism. Molybdenum is included in enzyme content which is called molybdoenzyms. Having bonded with molybdopterin it creates molybdenum co-factor (Moco) and gets oxidation-reduction properties. Moco is included in active site of molybdoenzymes. They take part in sulfur and nitrogen metabolism, and detrimental compound detoxication. Molybdenum deficiency is characterized by the slow plant growth, low amount of chlorophyll ascorbic acid capacity.It was noticed that plants suffering from the molybdenum deficiency can be saved, sodium molibdate can be used, it can be put directly in the soil or plant leaves can sprayed with the solution. There are five plant molibdoenzymes which are currently known: sulfite oxidase (SO), xanthine dehydrogenase (XD), nitrate reductase (NR), aldehyde oxidase (AO) and mitochondrial amidoxim-regenerative component. Nitrate reductase catalyzes the first stage of nitrate assimilation, eucariotic organisms contain three isoforms of the molybdoezimes: A NADH, A NAD(P)H и NADPH. Xanthine dehydrogenase regulates purine metabolism. XD increases plant antioxidant ability and slows down leaves aging. Molybdoenzymess are involved in the process of the stress adaptation, defining of the mechanisms and their reaction to environmental stress conditions is important for plant stress resistance.

Secretory Expression Of The Glucan Endo-1,3-Beta-D-Glucosidase Gene Of Secale Cereale In Yeast Pichia Pastoris

For survival in cold conditions, many organisms have developed unique adaptive mechanisms based on the synthesis of antifreeze proteins, peptides and glycoproteins that prevent ice formation at negative temperatures. These molecules tend to bind ice crystals and lower the freezing point of the solution without the formation of large crystals. Antifreeze proteins (AFP) were found in almost all types of living organisms, including insects, fungus, yeasts, bacteria and plants. The gene of antifreeze protein - glucan endo-1,3-beta-D-glucosidase (ScGlu-3) from Secale cereale was cloned into shuttle vector pPICZαA. The competent cells of yeast Pichia pastoris GS115 were transformed and the producer strain was obtained, which secreted of ScGlu-3 into the culture medium using 3% methanol as the only carbon source. It was found by western blotting that the maximum accumulation of ScGlu-3 in the culture occurs after 48 hours of fermentation on a medium with methanol. Established that rScGlu-3 precipitates at 50-65% of ammonium sulfate.

Obtaining Multiprotein Antigens Of Brucella And Study Of Their Immunoreactivity

Rose-Bengal test, complement fixation test and the agglutination test are mainly used for the diagnosis of animal brucellosis. These tests are characterized by low sensitivity and specificity, which is one of the main reasons for the low effectiveness of measures aimed at eradicating brucellosis. The use of modern highly sensitive serological tests requires the availability of antigens specific to Brucella spp. The aim of the study was to obtain multiproteins of the pathogen by recombinant DNA technology and to study their antigenic properties. In the course of the study, three types of multiproteins were obtained, constructed from diagnostically important peptides that form B.abortus and B.melitensis outer membrane proteins. All target products were synthesized by the producer strain in a form that is authentic to natural proteins, and showed immunogenicity in mouse model. Antibodies produced against the multiproteins were specific for the single proteins of the pathogen's cell wall. The data obtained indicate the need to continue studies to determine the possibility of using multiproteins as an antigen in an enzyme-linked immunosorbent assay to detect anti-Brucella specific antibodies.

Modification Of Taq DNA Polymerase with Improved Elongation Ability

Thermostable polymerases play a significant role in molecular biology and diagnostic practice. The most famous and demanded is Polymerase I from the thermophilic bacterium Thermus aquaticus (Taq-pol). This polymerase at one time made a kind of revolution in the polymerase chain reaction. In this work, we attempted to modify this polymerase by attaching an additional Sso7d protein from Sulfolobus solfataricus to Taq-pol, which provides additional binding to the double-stranded DNA of the template. Sso7d-Taq fusion gene was expressed in BL21(DE3) cells. Optimal conditions were selected for maximum production of modified Sso7d-Taq polymerase. The optimal conditions for the intracellular accumulation of Sso7d-Taq polymerase: activation of the T7 promoter when the optical density of the culture reaches OD600 = 0.8-1.0 by adding IPTG at a concentration of 0.2 mM, followed by incubation of the culture at 37°C for 20-24 hours. Recombinant Sso7d-Taq polymerase has been purified and tested by PCR for thermal stability and elongation time. It was found that the Sso7d-Taq enzyme withstands 5 hour incubation at 95°C and 75 minute incubation at 98°C. Comparative analysis with unmodified Taq DNA polymerase showed that the Sso7d-Taq enzyme reduces the elongation rate by several times - up to 15-13 seconds per 1 kbp. The results obtained indicate the prospects of using Sso7d-Taq DNA polymerase in scientific research and diagnostic practice.

Study Of Genetic Diversity And Resistance Of Fruit Crops To Main Pathogens Using DNA Markers

Breeding modern varieties of fruit crops requires the study of their biodiversity as a source of genes for useful traits, with the aim of transferring them to genome of commercial varieties. Application of genomic technologies can significantly speed up the breeding process. Identification and application of DNA markers for the study of genetic diversity, varietal identification, as well as the transfer of genes of valuable economic traits through marker assisted selection programs is of great applied importance. The article discusses the results of studies to identify genes and DNA markers associated with resistance of some fruit crops to major diseases.

INFLUENCE OF ABIOTIC STRESS FACTORS ON THE HYDRATION OF PLANT TISSUES OF SEDUM HYBRIDUM L. (AIZOPSIS HYBRIDA (L.) GRULICH)

Genus Sedum (family Crassulaceae) - succulents adapted to lack of moisture. Morphophysiological reactions of immature Sedum hybridum L. (Aizopsis hybrida (L.) Grulich) plants to stressful conditions of water scarcity, salinization and low positive temperatures are described. The high resistance of plants to the studied stress effects is shown. The tendency of the dynamics of the highest moisture loss by plants of the control group and the lowest by plants cultivated at PEG–6000 at a concentration of 200 mmol/l was noted, which indicates the adaptive effect of this level of osmotic stress on Sedum hybridum plants. To obtain a completely dry Sedum hybridum mass for various physiological experiments, it is necessary to maintain the plant material at a temperature of 105⸰ C, with at least 40 hours.

EXPRESSION OF THE STAPHYLOCOCCUS AUREUS LUKE GENE IN THE ESCHERICHIA COLI BL21(DE3) STRAIN

Two-component leukotoxins are important virulence factors for Staphylococcus aureus. Despite efforts made to study S. aureus leukotoxins, the direct mechanism of action of these toxins during infection has not been determined. However, the observation that deletion of LukED significantly attenuates highly virulent S. aureus strains supports the hypothesis that selective inhibition of LukE / D may be useful in the development of new aspects of S. aureus infection control. For this purpose, this work was carried out to test the expression and obtain a recombinant form of the LukE protein in E.coli cells. The LukE gene was cloned into the pET28-c (+) / GFP vector containing the gfp gene. Two fused genes carrying a hexahistidine tag were expressed in cells of the E. coli BL21(DE3) strain. It was found that the 6His-GFP-LucE protein aggregates in inclusion bodies. 6His-GFP-LucE was washed out of inclusion bodies with high molar urea. The 6His-GFP-LucE protein was purified by metal affinity chromatography. Research results can be applied to obtain recombinant protein including strategies for inhibition of toxin activity.

MICROCLONAL PROPAGATION OF SPIRAEA JAPONICA FOR LANDSCAPING

Japanese spiraea (Spiraea japonica) is an ornamental shrub widely used in landscaping. The method of clonal micropropagation of Spiraea japonica was optimized to obtain a large number of plants from several shoots. The optimal concentrations of hormones have been established to increase multiplication and root formation. QL medium with 0,5 mg/l benzyl aminopurine (BAP) in combination with 1,0 mg/l gibberelic acid (GA); 0,01 and 1,0 mg/l indolyl butyric acid (IBA); 1,0 mg/l of naphthyl acetic acid (NAA) were tested for multiplication. For root induction, naphthyl acetic acid (NAA) was used in five doses at half the concentration of QL and MS medium. The highest multiplication of shoots (14,02±1,39) and the highest increase in shoot length (6,39) was obtained on QL medium supplemented with 0,5 mg/l BAP; 1,0 mg/l GA and IBA 0,01 mg/l. The highest rooting (100%), the maximum number of roots (6,20±0,63), the length of the longest root (4,60±0,02) was observed on ½ QL medium containing 0,1 mg/l NAA. In conclusion, for Spiraea japonica, an efficient high speed and rooting protocol is described that can be used in mass propagation.

OPTIMIZATION OF PROCESS PARAMETERS FOR CULTIVATION OF BACTERIOPHAGES PROMISING FOR BIOLOGICAL CONTROL OF PHYTOPATHOGENIC BACTERIA OF GENUS PSEUDOMONAS

Cultural parameters of bacteriophages screened for lytic activity toward host bacterial strain P. helmanticensis BIM В-582 D were investigated. The following conditions were shown to be optimal for cultivation of indicator culture and it’s lysis by Pseudomonas phages BIM ВV-45 D, BIM ВV-46 D, BIM ВV-47 D, BIM ВV-50 D, BIM ВV-53 D, BIM ВV-61 D in laboratory fermenters ANKUM-3M: nutrient medium – fish meal hydrolysate broth, temperature – 28°C, stirrer agitation rate – 160 rpm, aeration rate – 1 l air/ l medium per min. Biopreparation produced under optimized conditions and composed of six afore-mentioned phages display high lytic activity both in regard to the host strain and to phytopathogenic bacteria Pseudomonas syringae.