[Analysis of galactose-1-phosphate uridyltransferase from different sources using electrophoresis in polyacrylamide gel].
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Animals↗
Biomedical subjects
T I Tikhonenko
Publications and source records attributed to T I Tikhonenko.
[Effect of nuclease S1 on DNA of the highly oncogenic simian adenovirus SA7(C8)].
The effect at specific nuclease S1 on DNA and the complex viral DNA-terminal protein of the highly oncogenic simian adenovirus SA7(C8) was studied. It was shown that nuclease S1 did not digest the bound between DNA and terminal protein in the complex but residual amino acid(s) was cleaved out after digestion with pronase. The DNA obtained after nuclease S1 action could be ligated and its 5'-ends were phosphorylated by polynucleotide kinase.
Adenoviridae↗
[Characteristics of integration of monkey adenovirus SA7 DNA fragments with genomes of transformed and tumoral cells].
Transformed rat cells and hamster tumoral cells, obtained after treatment with DNA of monkey adenovirus SA7 which was fragmented using endonuclease SalI, contained all the SalI fragments of SA7 DNA integrated with the cell genomes. As shown by blotting hybridization distinct deletion did not form in the fragments of viral DNA on integration. Insertion of the SA7 DNA fragments appears to occur into the definite relatively small site of the cell chromosome. In the both cell strains studied a site of the SA7 genome transcribed at coordinates: 1.8-10 units of the physical map. Fragments of viral DNA were methylated in the sites CmCGG and CCmCGGG of these both strains studied.
Adenoviridae↗
[Prospects and achievements of genetic engineering in development of antiviral vaccines].
Proceeding from the known data various theoretical and experimental approaches to the construction of gene-engineering vaccines are considered. Gene-engineering subunit vaccines of the first generation are based on isolation of the genes coding for the synthesis of full length capsid proteins with the main antigenic determinants and their subsequent expression in suitable recipient cells. Initial idea of the microbiological synthesis as the main way for production of any antiviral vaccines was not confirmed by the later development. Now for this type of vaccines eucaryotic systems are widely employed using the animal virus vectors and the animal cell cultures. Gene-engineering subunit vaccine of the second generation appears to be a chimeric protein with built-in antigenic determinants of different viruses and maximal immunogenicity in monomeric form. The last point reopens the perspective to use a microbiological synthesis for the production of antiviral vaccines. Besides that the chemically synthesized polypeptide antiviral vaccine will be used widely. In gene-engineering subunit vaccines of the third generation it is possible to use not the natural antigenic determinants which often are characterized by high level of the primary structure changes but artificial (non-natural) antigens, that are the capsid protein conservative regions which under natural conditions of infection or immunization do not induce the protective antiviral antibodies. The recombinant DNA technology in addition to subunit type vaccine allows to construct living vaccines which represent a DNA-containing attenuated virus with build-in natural or synthetic gene of the capsid or chimeric protein with antigenic determinants of another viral species.
Animals↗
[Optical rotatory dispersion by 5 viruses of the tobacco mosaic virus group and their components].
Optical rotatory dispersion (ORD) spectra in 250 to 350 nm region were measured for preparations of five TMV-like viruses (TMV vulgare, HR and U2 strains of TMV dolihosenation mosaic virus and cucumber virus 4) and also for RNA and protein preparations of these viruses. The data obtained testify against the possibility that the double peak with maxima at 286 and 293 nm observed in ORD of all the five viruses is due to interaction of tryptophan residues in virus coat protein with the RNA of the virul particle. The spectra of intravirus RNA of the five viruses, calculated as the difference between ORD of the intact virus and of its coat protein, were found to differ significantly from each other and from ORD of free RNA. ORD spectra of hybrid viruses, reconstituted from RNA of one virus and coat protein of another, proved to be identical to the ORD of the virus, whose protein was used in reconstitution. We suppose that the difference in ORD of the intravirus RNA of the five viruses reflect differences of RNA-protein interactions in them.
Chemical Phenomena↗
[The problem of classifying simian adenoviruses. Electrophoretic and immunologic analysis of hexons].
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Adenoviridae↗
[Comparative study of variants of the Venezuelan equine encephalomyelitis virus isolated from chronically infected cells by transfection in different cells].
Comparative data with regard to the properties of Venezuelan equine encephalomyelitis (VEE) virus isolated from HeLa carrier cultures by transfection in different cell cultures have been obtained. Introduction of DNA extracted from the carrier cultures into BHK-21 cell cultures resulted in production of an actively multiplying medium-plaque virus, and parallel addition of the same DNA preparations in chick fibroblast or monkey kidney cultures led to production of small-plaque virus with a low reproduction potential. The virus produced by transfection of BHK-21 cells differed from that produced in chick fibroblast and monkey kidney cultures in electrophoretic mobility of virion envelope proteins. The infection of these cultures with virions as well as infection with genome RNA did not result in production of differing virus variants. The importance of the experimental genetic data for the problem of the nature of the infectious principle of cellular DNA preparations and of the form of existence of viral genome in chronically infected cultures with infection of the integrative type is discussed.
Animals↗
[Electron microscope maps of SA7 DNA melting].
Electron microscopic denaturation maps corresponding to the first peaks of the differential melting curve of SA7 DNA were constructed by fixation of partly denatured molecules with glyoxal at temperatures within the melting range. These maps were oriented with respect to the functional map of the virus genome. The localization and the size of the most AT-rich SA7 DNA regions were determined.
Adenoviridae↗
[Calculating light scattering when determining "true absorption magnitudes" of virus particles].
Results of the study by V. I. Permogorov et al. (Molecular Biology, 1977, 11, 134--138) on the absence of a deficit of hypochromism in intraphage DNA are discussed. It is deduced that the conclusion of V. I. Permogorov and coworkers is erroneous since it is based on: 1) incorrect interpretation of the results of determination of lightscattering contribution to the absorption of intact phages; 2) not taking into account the lightscattering contribution in the case of disrupted phages; 3) underestimation of the value of hyperchromism of melting of phage DNAs.
DNA, Viral↗
[Some results of research according to the "gene" program].
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Cloning, Molecular↗
[Bacteriophage P22 DNA structure in situ].
The structure of phage P22 DNA in situ was investigated by optical methods and by chemical modification with sodium bisulfite. On disruption of the phage particles by heating at 45 degrees a drop in absorbance at the 250 nm to 290 nm region was observed. At 260 nm this hypochromism was about 12%. CD spectra of intraphage DNA differed from that of free P22 DNA in the intensity as well as in the position of the positive band (lambda max 280 nm, delta epsilon max=1.3). In the intraphage DNA 21 per cent of cytosines reacted with sodium bisulfite. Cytosyl-amino acid products were found in the HClO4 and HCl hydrolysates of the modified phage. The main amino acid component of the product was identified as lysine. It was shown by means of gradient centrifugation and electron microscopy that the cytosyl-amino acid products result in the crosslinking of DNA to protein in the phage particles.
Circular Dichroism↗
[Synthesis of adenovirus SA7 and Ad5 DNA in extracts from the nuclei of green monkey kidney cells].
A soluble extract from the nuclei of green monkey kidney cells, infected with adenovirus SA7, carries out replication of SA7 DNA. It was observed, that in vitro DNA synthesis, which is dependent on the exogenously added viral DNA-protein complex as its optimal template, proceeds by a semiconservative mechanism. The data obtained are compared with literary data on soluble enzyme systems from adenovirus-infected cells. The early factors provided by SA7 also supported replication of the DNA-protein complex, prepared from human adenovirus type 5, in nuclear extracts of monkey cells.
Adenoviruses, Human↗
[Current biotechnology: its limits and prospectives].
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Biochemistry↗
[Isolation and translation of late mRNA coding for the structural proteins of simian adenovirus SA7].
A total preparation of cytoplasmic RNA was isolated from late stage-infected African green monkey kidney cells using phenol extraction procedure. The poly (A)-containing mRNA fraction was selected on oligo(dT)-cellulose columns. The resulting mRNA preparations were heterogenous in size and contained about 20--60% of SA7-derived sequences. SA7 late mRNA was efficiently translated in rabbit reticulocyte cell-free system giving rise to a number of polypeptide products that were related by antigenicity to authentic SA7 virion proteins. The main translation product having a molecular weight of 114 kilodaltons was identified as intact SA7 hexon protein.
Adenoviridae↗
[Identification of proteins and amino acids reacting with DNA in the composition of bacteriophage P22].
An interaction occurring between DNA cytosine and phage P22 protein was fixed using chemical modification with a nucleophilic agent, sodium bisulphite. Four minor proteins (P1, P20, P26, and P4) interacting with phage P22 DNA were isolated. Amino acids lysine and arginine were found to be the immediate partners of cytosine realizing this interaction.
Amino Acids↗
[Physical characteristics of simian adenovirus SA7].
Physical characteristics of simian adenovirus SA7 virion were studied. The buoyant density of SA7 virions was found to be 1.356 g/cm3. The percentage of DNA calculated from the buoyant density value and determined with phosphorus directly was 11.6% and 11.8%, respectively. The S020,w and D020,w were 740 S and 3.34 x 10(-8)/cm2, respectively. From the hydrodynamic parameters of the virion and its volume and specific partial volume as well as percentage of DNA and DNA molecular weight, the molecular weight of SA7 virion was calculated and found to be 185.5 Mdaltons.
Adenoviridae↗
[Infectivity of the DNA of PM-2 phage].
The infectivity of PM-2 phage free DNA was studied versus competent cells of marine bacteria Pseudomonas Bal-31 depending on the time of incubation with the bacterial cells, on the DNA and calcium concentration in the solution. Optimal conditions for the infection are the following: DNA concentration about 3 microgram/ml, CaCl2 concentration 0.1 M, incubation time 20 min.
Bacteriophages↗
[Fragmentation of human adenovirus type 1 DNA by the specific endonucleases R. SaLI, R. EcoRI and R. BamHI].
The restricting endonuclease R. EcoRI hydrolyses human adenovirus type I (Adl) DNA with formation of three fragments with molecular weights of 17.5 x 10(6), 3.7 x 10(6), and 1.55 x 10(6) dalton. Under the effect of R. SaLI restrictase DNA Adl disintegrates into 5 fragments with molecular weights: 8.0 x 10(6), 6.0 x 10(6), 4.6 x 10(6), 4.0 x 10(6), and 0.2 x 10(6) dalton. Treatment of DNA with R. BamHI produces 4 fragments with molecular weights 10.0 x 10(6), 6.8 x 10(6), 4.0 x 10(6), and 2.2 x 10(6) dalton. Fragments of R. SaLI and R. BamHI form sequences BECAD and ACBD, respectively.
Adenoviruses, Human↗