[Expression of cecropin P1 gene increases resistance of Camelina sativa (L.) plants to microbial phytopathogenes].

Transgenic plants of camelina (Camelina sativa (L.) Crantz) with the synthetic gene of antimicrobial peptide cecropin P1 (cecP1) were obtained. Agrobacterium-mediated transformation is performed using the binary vector pGA482::cecP1 by vacuum infiltration of flower buds.

[Peculiarities of 5-methylcytosine distribution in eukaryotic genome].

Most of it functions DNA methylation realizes as an integral part of the mechanism of remodeling and modification of chromatin structure. At the same time the global pattern of this complex reaction's net is still to be determined and we are just approaching to studying the mechanisms controlling epigenetic processes of histone modification and DNA methylation. Though cytosine methylation occurs predominantly at CpG sequences of eukaryotic genome, it also takes place at symmetric CpHpG and non-symmetric CpHpH sites (where H-A, T, or C).

[The use of RNA interference for the metabolic engineering of plants].

The metabolic engineering of plants is aimed at the realization of new biochemical reactions by transgenic cells. These reactions are determined by enzymes encoded by foreign or self-modified genes. Plants are considered to be the most interesting objects for metabolic engineering.

[The influence of colonizing methylobacteria on morphogenesis and resistance of sugar beet and white cabbage plants to Erwinia carotovora].

The influence of colonization of sugar beet (Beta vulgaris var. saccharifera (Alef) Krass) and white cabbage (Brassica oleracea var. capitata L.

[Inhibition of agrobacterial oncogenes expression by means of antisense RNA].

Plant's infection with soil bacteria Agrobacterium tumefaciens lead to tumour formation, so called crown galls. The reason of tumorigenesis is integration of agrobacterial genes for phytohormone synthesis auxins and cytokinins in plant genome, the most important of them are iaaM and ipt. Obtaining of transgenic plants able to inhibit these genes expression, creates conditions for producing of plants resistant to crown gall disease.

[Enhanced resistance to phytopathogenic bacteria in transgenic tobacco plants with synthetic gene of antimicrobial peptide cecropin P1].

Plasmids with a synthetic gene of the mammalian antimicrobial peptide cecropin P1 (cecP1) controlled by the constitutive promoter 35S RNA of cauliflower mosaic virus were constructed. Agrobacterial transformation of tobacco plants was conducted using the obtained recombinant binary vector. The presence of gene cecP1 in the plant genome was confirmed by PCR.

[Biosynthesis of mini-antibodies to human ferritin in plant and bacterial producers].

A recombinant scFv antibody against human spleen ferritin was expressed as a barstar-fused protein in Escherichia coli and in Nicotiana tabacum plants and suspension cell cultures. As demonstrated by immunoblotting with antibarstar antibodies, direction of the recombinant protein to the endomembrane system of plant cells ensured its stability and solubility. Production of the recombinant protein did not differ between parental transgenic plants and their first-generation progeny.

[Analysis of transgenic tobacco plants carrying the gene for the surface antigen of the hepatitis B virus].

The plasmids carrying the gene encoding the hepatitis B surface antigen (HBsAg) under the control of 35S RNA single or dual promoters of the cauliflower mosaic virus CaMV 35S were constructed. These constructions were used for obtaining transgenic tobacco plants that synthesize the HBS antigen. The presence of HBsAg in tobacco plant extracts was confirmed by the enzyme-linked immunoassay using antibodies against the native HBs antigen.

[Hypermethylation of the human calcitonin gene as a molecular marker in acute lymphoid leukemia].

HpaII/MspI blot-hybridization analysis of the 5'-end region of the calcitonin (CT) gene methylation in cells of bone marrow and peripheral blood of patients with acute lymphoblastic leukemias (ALL) has been carried out. ALLs are accompanied by hypermethylation of the inner cytosine in the CCGG sequences of this region of the CT gene. The level of hypermethylation of the CT gene corresponded to the degree of disease progression and malignancy.

[Phenotypic changes in transgenic tobacco plants with an antisense form of the hmg1 gene].

Tobacco plants were genetically transformed with the Arabidopsis thaliana heterologous hmg1 gene encoding 3-hydroxy-3-methylglutaryl-CoA reductase, a key enzyme involved in the metabolism of terpenoid compounds. The hmg1 gene was inserted under the control of the 35S RNA double promoter from the cauliflower mosaic virus (CaMV 35S) both in direct and reverse orientation relative to the promoter. DNA analysis by polymerase chain reaction (PCR) and Southern blotting confirmed the transgenic nature of the tobacco plants obtained.

[The lack of the CpNpG methylation at the 5'-terminal region of the human calcitonin gene in norm and in leukemia [].

The inner cytosine methylation was analyzed in the CCWGG sequences of the 5'-terminal region of the human calcitonin gene from peripheral blood and bone marrow cells in various forms of leukemia. Since these sequences remain nonmethylated both in norm and in various leukemia forms, the CpG dinucleotide hypermethylation of the 5'-terminus of the human calcitonin gene, characteristic for the development of leukemias, does not spread over adjacent CpNpG sequences.

[DNA methyltransferases for detection of the level of methylation of cytosine in the DNA CCWGG sequence].

An assay for the cytosine methylation level in the eukaryotic DNA CCWGG sequence is proposed. The method is based on the ability of DNA methylase BstNI to methylate DNA containing in a CCWGG site a nonmodified or 5-methylated cytosine to yield N4-methyl- or N4,5'-dimethylcytosine, respectively.

[Formation of deletional derivatives of the Ti-plasmid pGV3850 in a conjugative transfer from Agrobacterium tumefaciens to Escherichia coli].

The process of the transfer of the Ti-plasmid vector pGV3850 with the plasmid pBR322 inserted into the T-DNA region from Agrobacterium tumefaciens to a non-plasmid strain of Escherichia coli was studied. The transferred Ti-plasmid was found to be exposed to deletions and formed a wide range of derivatives with a size ranging from 3-4 kb to 50 kb, maintained in E. coli due to ColE1-replicon.

[Formation of two types of enzyme-substrate complexes during the interaction of EcoRII restriction enzyme with synthetic DNA-duplexes].

Binding of EcoRII restriction endonuclease to synthetic oligodeoxyribonucleotide substrates of 11-30 base pairs long was investigated by polyacrylamide gel electrophoresis under nondenaturing conditions in the absence of Mg2+ ions. Irrespective of the length of a substrate, two types of specific DNA-protein complexes were shown to be formed. Their mobility in gel was close to that of the monomer (45 kDa) and dimer (90 kDa) of marker protein, ovalbumin.

[Interaction of EcoRII restriction and modification enzymes with synthetic DNA fragments. Determination of the size of EcoRII binding site].

EcoRII restriction endonuclease binding site has been determined on the basis of comparison of the binding parameters of the enzyme with synthetic DNA-duplexes of concatemer type containing a different number of EcoRII recognition sites. It has been shown that it consists of 21 +/- 1 base pairs.

[UV- and CD-spectra of restriction endonuclease EcoRII and DNA-methylase EcoRII].

UV- and CD-spectra of homogeneous enzymes have been measured. Extinction coefficients estimated from the UV-spectra are 0.97 for restriction endonuclease EcoRII at 279.

[Isolation, purification and properties of restrictase and methylase BstN1 from Bacillus stearothermophilus].

Restriction-methylation enzymes BstN1 from Bacillus stearothermophilus were isolated and purified. These enzymes are related to a new class of restriction-methylation enzymes of the second type, whose modifying component is N4-cytosine-DNA-methylase. Both enzymes recognize the DNA sequence CC(A/T)GG.

[Purification of restriction endonuclease EcoRII using monoclonal antibodies].

Monoclonal antibodies against EcoRII endonuclease were obtained after immunization of two BALB/c mice with a homogeneous enzyme prepared by conventional methods. IgG from ascitic fluid was purified and coupled to CNBr-activated Sepharose 4B to give a specific column used to isolate EcoRII endonuclease. The isolated EcoRII endonuclease produced a single band during SDS gel electrophoresis.

[A method for S-adenosylmethionine determination in extracts of microbial cells].

A technique for quantification of S-adenosylmethionine in microbial cell-free extracts is proposed that involves dilution of S-adenosyl-L-(methyl-3H)methionine with non-labelled S-adenosylmethionine followed by DNA-cytosine-methyltransferase reaction. The content of S-adenosylmethionine and the activity of S-adenosylmethionine synthetase in yeasts and E. coli MRE-600 are in good agreement with the results obtained with labelled L-methionine and consistent with literature data.

[A method for detecting new strains producing DNA modification and restriction enzymes--isoschizomers and isomethylomers of known restrictases and methylases].

The paper describes a technique for the detection of new strains producing enzymes which mediate DNA modification and restriction, and isoschizomers and isomethylomers of the known restriction endonucleases and methylases. Three Bacillus subtilis strains whose DNA carries a BamH1 modification have been found. Two of these strains exert the restrictase activity with an R BamH1 specificity.

[Isolation of modification-restriction enzymes HpaI and HpaII].

A method for simultaneous isolation of four enzymes of modification-restriction of DNA from Haemophilus parainfluenzae is proposed. The properties of HpaI and HpaII DNA-methylases were investigated.

[Effect of Ecodam DNA-methylase on single-stranded sequences and synthetic oligonucleotides].

Interaction of the Ecodam methylase with different substrates were investigated among them the double- and single-stranded DNAs and synthetic oligonucleotides containing some defects in the GATC sequence. These defects were:nick, the absence of one internucleotide phosphate of nucleotide; partially single-stranded form on the recognition site etc. It was demonstrated that the presence of both G .