Immunogenicity and Protective Efficacy of Influenza A DNA Vaccines Encoding Artificial Antigens Based on Conservative Hemagglutinin Stem Region and M2 Protein in Mice.

Background: Development of a universal vaccine capable to induce antibody responses against a broad range of influenza virus strains attracts growing attention. Hemagglutinin stem and the exposed fragment of influenza virus M2 protein are promising targets for induction of cross-protective humoral and cell-mediated response, since they contain conservative epitopes capable to induce antibodies and cytotoxic T lymphocytes (CTLs) to a wide range of influenza virus subtypes.

Methods: In this study, we generated DNA vaccine constructs encoding artificial antigens AgH1, AgH3, and AgM2 designed on the basis of conservative hemagglutinin stem fragments of two influenza A virus subtypes, H1N1 and H3N2, and conservative M2 protein, and evaluate their immunogenicity and protective efficacy.

Attenuated Salmonella enteritidis E23 as a vehicle for the rectal delivery of DNA vaccine coding for HIV-1 polyepitope CTL immunogen.

This study is focusing on elucidation of the capacity of attenuated Salmonella enteritidis E23 (cya, crp) to serve as a vehicle for the rectal delivery of the DNA vaccine. Earlier for creation HIV-1 candidate DNA vaccine we have designed the polyepitope protein TCI (T-cell immunogen), which comprises over 80 CTL epitopes from subtype A, B and C HIV-1 proteins. The gene coding for TCI protein was used to construct the eukaryotic expression plasmid pcDNA-TCI.

[Mechanisms of HIV-1 drug resistance to nucleoside and non-nucleoside reverse transcriptase inhibitors].

Global AIDS epidemics caused by human immunodeficiency virus type 1 (HIV-1) has existed for more than 25 years and involved more than 2 million newly infected people annually. The obstacle in combating the global epidemics is a rapid evolution of the virus by the selection of drug resistance mutations. In this review, we have summarized scientific achievements in the field of reverse transcriptase drug resistance to licensed antiviral drugs--nucleoside (NRTI) and non-nucleoside (NNRTI) inhibitors.

[RT-PCR-based methods for identification and typing of infectious hemopoietic necrosis virus in salmons].

A RT-PCR method has been developed to diagnose infectious hemopoietic necrosis virus (IHNV) in salmons. The authors show it possible to use the method for viral shedding in both a cell culture and a clinical sample from infected fishes. Genotyping of IHNV strains originating from North America, Europe, and Russia, by using the restriction fragment length polymerase analysis, has revealed that 10 of them belong to 3 existing genogroups (U, M, and L).

[Genomic and phenotypic analyses of microorganisms isolated from the sediments of Lake Baikal].

Genetic and biochemical methods and morphological examination were used to study microorganisms isolated from samples of deep drilling of the Lake Baikal bottom sediments. Based on blot hybridization patterns, the strains investigated were divided into several groups according to the degree of homology of their genomic DNA. Morphological, biochemical, and ultrastructural characteristics of bacterial strains are described, and their compliance with the genomic analysis data is demonstrated.

[Genetic polymorphism of clinical Mycobacterium tuberculosis strains circulating in the territory of Novosibirsk Region].

Mycobacterium tuberculosis strains isolated from patients treated at TB dispensary branches in different districts of Novosibirsk were studied by genetic analysis. The below molecular methods were used: 1. PCR with random primers; 2.

[DNA technology for diagnosis and characterization of agricultural animal pathogenic viruses].

Use of recombinant DNA for the development of diagnostic and therapeutic and preventive drugs became one of the priority trends in modern experimental veterinary. This paper discusses modern methods of virus analysis based on the DNA technologies: restriction mapping, nucleic acid hybridization, and polymerase chain reaction. Examples of utilization of these methods for clinical diagnosis and research of animal viruses are offered.

[Structure of over lapping region of ICP18.5 and gB genes of type 1 bovine herpesvirus strain TK-A].

Nucleotide sequence of DNA fragment coding 3'-terminal of ICO 18.5 gene that overlaps the regulatory region and 5'-terminal of open reading frame of gB gene of bovine herpesvirus (BHV-1, subtype 1.3), strain TK-A, was determined.

Detection of spring viremia of carp virus isolates by hybridization with non-radioactive probes and amplification by polymerase chain reaction.

For detection of spring viremia of carp virus (SVCV) DNA probes have been constructed using the reverse transcription-polymerase chain reaction (RT-PCR) amplification technique and cDNA cloning in plasmid and phage vectors. The specific primers for amplification of SVCV M and G genes were chosen and synthesized. Studies were carried out to establish the sensitivity and specificity of viral RNA detection in infected cell culture and pathogenic material from fish by the use of non-radioactive probes and RT-PCR.

[Identification and characterization of strains of Bacillus thuringiensis by genomic fingerprinting using biotinylated phage M13 DNA].

Genomic DNA of the entomopathogenic bacterium Bacillus thuringiensis was analyzed by the genomic fingerprinting technique. The biotin-labeled single-stranded DNA of the phage M13 was used as a marker of hypervariable sequences. A procedure for analyzing the differentiation among various Bacillus thuringiensis strains was developed.

[Detection of the hog cholera virus using the polymerase chain reaction].

A rapid and highly sensitive method for detecting hog cholera virus (HCV) based on a reverse transcription of the polymerase chain reaction (RT-PCR) is developed. Primers complementary to the most homologous sites of virus genome in an area coding the precursor for glycoproteins gp44/gp48 are selected. Detection of the virus in pathological material by the RT-PCR showed that use of these primers in amplification allows detection of different HCV strains.

[Identification and classification of strains of microorganisms using genomic fingerprinting with biotinylated phage M13 DNA].

To analyze DNA polymorphisms of various bacterial strains, a nonradioactive variant of the genomic fingerprinting method was developed. The method was based on the application of biotin-labeled single-stranded phage M13 DNA as a probe. Characteristic patterns of fingerprints obtained by MvaI, HaeIII, and HinfI restriction enzymes are presented for several species of bacilli and other bacteria.

[Detection of the spring viremia of carp virus by hybridization with nonradioactive DNA probes].

In order to detect spring viraemia of carp virus, DNA probes have been constructed using reverse transcription-PCR amplification technique and cDNA cloning in plasmid and phage vectors. The sensitivity and specificity of viral RNA detection was assessed using nonradioactive probes in infected FHM cell culture and in tissues from dead fish. Viral RNA was more frequently detected in the brain and gill than in the abdominal organs.

[Detection of bovine infectious rhinotracheitis virus by hybridization using nonradioactive DNA-probes].

Biotin-labeled DNA probes for infectious bovine rhinotracheitis virus also known as bovine herpesvirus-1 (BHV-1) have been developed. The procedure is based on dot-blot hybridization using biotin-labeled bacteriophage M13 and plasmid probes containing cloned PstI and EcoRI-PstI restriction fragments of viral genome. The probes obtained were used to detect viral nucleic acids in specimens of bovine spermatic fluid or nasal swabs of calves.

[Preparation of a specific immunogen based on bacteriophage M13].

Earlier we developed an expression vector on the basis of bacteriophage M13 allowing the exposure of short peptides on the virion surface. It was used to obtain a recombinant phage carrying the antigenic determinant of HIVI gag protein p17. This phage was tested as immunogen in rabbits.

[Use of non-radioactive biotin-labelled probes for detecting hepatitis A virus].

The biotin-labeled DNA probes were constructed on the basis of the hybrid bacteriophage M13nip 9 single-stranded DNA containing the fragments of the hepatitis A viral cDNA. The probes were biotin treated by chemical modification of the DNA by the peraminating reagent or photochemically. The labeled DNA probes were used in molecular hybridization experiments with the nuclear acids fixed on the nitrocellulose filters.

[Behavior of the large fragment of DNA polymerase I (the Klenow fragment) during fractionation of a cell-free extract of E. coli MRE-600].

Distribution of the DNA polymerase I large fragment (Klenow fragment) was studied during fractionation of the E. coli MRE-600 cell-free extract with polyethylenimine. On the basis of the results obtained a simple procedure is proposed that enables the Klenow fragment to be obtained as a coproduct of DNA polymerase I, RNA polymerase, polynucleotide phosphorylase, nucleotide kinases with acetokinase and nucleoside deoxy-ribosyltransferase in the framework of a combined technological scheme.

[Isolation and properties of immobilized alkaline phosphatase from E. coli].

Preparations of alkaline phosphatase from E. coli, immobilized on Sepharose, with a specific activity of 40-60 U/g wet weight were obtained. The immobilized enzyme was stable up to 50 degrees C; at higher temperatures it was inactivated.

[Properties of phosphodiesterase from Vipera lebetina venom immobilized on Agarose].

Preparations of phosphodiesterase from Vipera lebetina venom immobilized on agarose were obtained. The kinetic properties for the hydrolyses of various substrates of soluble and immobilized phosphodiesterase, e. g.

[Preparation and study of properties of immobilized phosphodiesterases from blunt-nosed viper venom].

From gurza poison phosphodiesterase covalently bound with DEAE-cellulose via triazine dye was isolated. Immobilized phosphodiesterase retained 100% activity of the initial enzyme. The effect of pH, temperature.