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Biomedical subjects

P A Belavin

Publications and source records attributed to P A Belavin.

13 recordsLinked to original sources

[Different systems of delivery of HIV-1 DNA vaccine encoding the multiepitope CTL-immunogene].

We used, within the case study, virus-like particles (VLP) and attenuated strains of salmonella for the delivery of HIV-1 DNA vaccine encoding the multiepitope CTL-immunogene. The immunogenicity of the thus obtained vaccine constructions was comparatively analyzed. All constructions were shown to be able of inducing, in immunized animals, both the specific T-cell responses and the synthesis of virus-specific antibodies. The lowest level of immune response was registered in animals immunized by "naked" plasmid DNA. The delivery by plasmid DNA involving VLP or the attenuated strain of salmonella enhances the efficiency of the DNA-vaccine presentation to the immune system.

AIDS Vaccines↗

[Recombinant vaccinia virus expressing Japanese encephalitis virus protein E].

Recombinant vaccinia virus expressing protein E of Japanese encephalitis virus has been constructed. Polyclonal antibodies to JE virus reacted with recombinant protein E in immunoblotting. Immunochemical analysis of the recombinant protein E with monoclonal antibodies showed that both group specific and receptor domains of the protein were intact.

Cell Line↗

[Preparation of single-chained antibodies to surface glycoprotein E from tick-borne encephalitis virus].

A single-stranded antibody gene scFv was designed on the base of cDNA fragments of genes coding for variable domains of heavy and light chains of MAb E6B to tick-borne encephalitis glycoprotein E. High production of stable soluble scFv was reproduced in Escherichia coli cells. Recombinant antibodies bound to antiidiotypical antibodies to initial MAb E6B and to recombinant virus protein E. Competitive analysis showed that single-stranded antibodies inhibited reaction between MAb E6B and protein E. These results confirm the formation of scFv with the original antigen-binding specificity towards tick-borne encephalitis virus glycoprotein E.

Antibodies↗

[Interaction of tick-borne encephalitis virus RNA with proteins].

Binding of tick-borne encephalitis (TBE) virus RNA to proteins was studied by 3 methods: gel retardation, RNA-protein cross-linking under UV light, and RNA-protein blotting. Two cellular proteins with molecular weights about 30 and 22 kDa were detected in the nuclear fraction of two cell strains (PS and RH) and in blood mononuclear cells of patients with TBE. Weak interaction between the studied RNA and viral proteins can result from trace amounts of TBE virus proteins in infected cells and by lack of additional binding factors. Bacterial super-products and affinity chromatography for isolation of native glycoproteins were used to increase the amount of viral proteins. Purified E and NS1 proteins did not react with viral RNA, while incubation with recombinant nonstructural TBE virus NS3 protein led to a shift in the mobility of TBE virus RNA in gel in the absence of cross-linking under conditions of UV exposure after addition of heparin. Poor binding of nonstructural TBE virus NS3 protein is based on electrostatic binding. Oligoribonucleotide 14 nucleotide residues long corresponding to 5'-terminal of TBE virus genome did not react with NS3 protein, probably because of its small size or absence of certain sequences.

Animals↗

[Expression of orthopoxvirus DNA sequences in Escherichia coli cells].

We observed the expression of recombinant plasmids genes containing ectromelia virus DNA fragments in E. coli minicells. Using plasmids with vaccinia or ectromelia viruses DNA fragments inserted upstream of lacZ gene we showed that certain orthopoxvirus genome fragments carry out a promoter-like function in bacterial cells.

Autoradiography↗

[A method for detecting restriction endonucleases in bacterial colonies].

A simple technique is proposed for detection of bacterial restriction endonucleases. Analysis is performed directly in the cells from colonies cultivated on Petri dishes. The cells collected with an inoculation loop are treated with lysozyme and Triton X-100. After centrifugation the supernatant is tested for endonuclease activity. The technique enables up to 100 colonies to be tested for 3-4 h.

Bacteria↗

[Cloning and primary structure determination of genes coding for IE2 and PP150 proteins of human cytomegalovirus].

The epidemiological situation with cytomegalovirus infection in Siberia is still to be studied and serological diagnosis of human cytomegalovirus (HCMV) is not satisfactory. Two regions of HCMV genome (strain AD-169) have been examined for obtaining diagnostic reagents by expression cloning. Using polymerase chain reaction, fragments of gene of the immediate early protein (IE2) and of the region encoding for the entire hydrophilic part (1176 bp) of the large phospoprotein gene (pp150) have been obtained. Both fragments were cloned in bacterial vectors. Analysis of nucleotide sequence showed negligible substitutions in comparison with previously reported sequences for these genes.

Amino Acid Sequence↗