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10 results on '"Totmenin Av"'

1. [Expression of genes for orthopoxviral TNF-binding proteins and study resulted recombinant proteins].

Author

Gileva IP, Riazankin IA, Nepomniashchikh TS, Totmenin AV, Maksiutov ZA, Lebedev LR, Afinogenova GN, Pustoshiliva NM, and Shchelkunov SN

Subjects
Animals, Base Sequence, Cell Line, DNA Primers, Enzyme-Linked Immunosorbent Assay, Mice, Recombinant Proteins genetics, Recombinant Proteins metabolism, Spodoptera, Tumor Necrosis Factor-alpha toxicity, Viral Proteins metabolism, Orthopoxvirus genetics, Tumor Necrosis Factor-alpha metabolism, Viral Proteins genetics
Abstract

Genes for TNF-binding proteins (CrmBs) of variola virus (VARV), monkeypox virus (MPXV) or cowpox (CPXV) were isolated with PCR from viral genomes and expressed within baculovirus DNAs in Sf21 insect cell line. Properties of resulted recombinant proteins were studied with physical-chemical and immunological methods. It was shown with solid phase enzyme-linked immunoassay that viral proteins inhibited hTNF binding with polyclonal hTNF-antibodies. The strongest inhibitor was VARV-CrmB, the less one was MPXV-CrmB. Biological activity of recombinant protein preparations was studied in the test of neutralization of TNF cytotoxicity for L929 murine fibroblast cells. It was shown that recombinant CrmBs neutralized cytotoxicity of hTNF, mTNF or rTNF in species-specific manner. It was shown also that effectiveness of hTNF cytotoxicity inhibition in vitro with VARV-CrmB exceeded the same effect of polyclonal hTNF-antibody. A possibility of the elaboration of new therapeutics for anti-TNF therapy on the base of CrmB-like proteins is discussed.

Published
2005

2. [Orthopoxvirus genes for Kelch-like proteins. I. Analysis of species specific differences by gene structure and organization].

Author

Totmenin AV, Kolosova IV, and Shchelkunov SN

Subjects
Amino Acid Motifs, Amino Acid Sequence, Animals, Carrier Proteins metabolism, Gene Order, Genome, Viral, Humans, Molecular Sequence Data, Mutation, Orthopoxvirus pathogenicity, Phylogeny, Sequence Alignment methods, Sequence Homology, Amino Acid, Species Specificity, Viral Proteins metabolism, Carrier Proteins genetics, Orthopoxvirus genetics, Viral Proteins genetics
Abstract

Genes and proteins of the kelch superfamily were structurally analyzed in the smallpox (SPV), monkeypox (MPV), cowpox (CPV), and vaccinia (VV) viruses. Genes potentially coding for the kelch-like proteins were found only in the variable terminal regions of the orthopoxvirus genome. The set and sizes of their protein products varied with species. All genes of the superfamily proved to be disrupted by mutations in SPV, which is highly pathogenic for its only host, man. The largest set of kelch-like proteins was observed for CPV, which is low-pathogenic for humans and has the broadest animal host range. The kelch-like proteins of one virus showed low homology to each other, whereas isologs of different viruses were highly homologous. The results testified to the earlier assumption that CPV is the most ancient and an ancestor of the other orthopoxviruses pathogenic for humans.

Published
2002

5. [Study of the structure-activity organization of the smallpox viral genome. V. Sequencing and analysis of the nucleotide sequence of the left terminus of the India-1967 strain genome].

Author

Shchelkunov SN, Totmenin AV, Babkin IV, Safronov PF, Gutorov VV, Pozdnaikov SG, Blinov VM, Resenchuk SM, and Sandakhchiev LS

Subjects
Amino Acid Sequence, Animals, Base Sequence, Cells, Cultured, DNA, Viral, Molecular Sequence Data, Restriction Mapping, Sequence Homology, Amino Acid, Species Specificity, Structure-Activity Relationship, Viral Proteins genetics, Genome, Viral, Variola virus genetics
Published
1996

6. [Study of the structure-function organization of the variola virus genome. IV. Sequencing and analysis of the nucleotide sequence of the right terminus of the India-1967 strain genome].

Author

Blinov VM, Totmenin AV, Resenchuk SM, Olenina LV, Chizhikov VE, Kolykhalov AA, Frolov IV, Gutorov VV, Pozdniakov SG, and Krasnykh VN

Subjects
Amino Acid Sequence, Animals, DNA Ligases genetics, DNA, Viral, Molecular Sequence Data, Open Reading Frames, Plasmids, Receptors, Cytokine genetics, Sequence Homology, Amino Acid, Species Specificity, Structure-Activity Relationship, Thymidine Kinase genetics, Genome, Viral, Variola virus genetics
Abstract

Sequencing and computer analysis of the variola major virus strain India-1967 (VAR-IND) genome segment (53,018 bp) from the right terminal region have been carried out. Fifty nine potential open reading frames (ORFs) of over 60 amino acid residues have been identified. Structure-function organization of VAR-IND DNA segment under study was compared with the previously reported sequences from the analogous genomic regions of vaccinia virus strains Copenhagen (VAC-COP) and Western Reserve (VAC-WR) and variola virus strain Harvey (VAR-HAR). Multiple distinctions in the genetic map of VAR-IND from VAC-COP and VAC-WR have been revealed along with the high similarity to the corresponding VAR-HAR segment. Possible functions of the predicted viral proteins and the effect of their differences on the features of orthopoxviruses are discussed.

Published
1995

7. [Structure-activity organization of the variola virus genome. III. Sequencing and analysis of the nucleotide sequence of the conserved region of HindIII-F, -N-, and -A-fragments of the India 1967 strain genome].

Author

Shchelkunov SN, Resenchuk SM, Totmenin AV, Kolykhalov AA, Frolov IV, Dryga SM, Volchkov VV, Chizhikov VE, Gutorov VV, and Blinov VM

Subjects
Amino Acid Sequence, Base Sequence, DNA-Directed RNA Polymerases metabolism, Deoxyribonuclease HindIII, Genome, Viral, Inclusion Bodies, Viral, Molecular Sequence Data, Open Reading Frames, RNA, Viral genetics, Restriction Mapping, Transcription Factors metabolism, Viral Proteins genetics, Virion, Conserved Sequence, Variola virus genetics
Abstract

Computer analysis of variola major virus (VAR) genomic fragment bounded by open reading frames (ORFs) D1R and A33L which is 47,961 bp long revealed 46 potential ORFs. The VAR proteins were compared with the analogous proteins of vaccinia virus strain Copenhagen. The subunits of DNA-dependent RNA polymerase, as well as the transcription factors, mRNA capping enzymes, and proteins necessary for the virion morphogenesis proved to be highly conservative within orthopoxviruses. The most pronounced differences between the VAR genome fragment under study and the corresponding vaccinia virus fragment were revealed in the vicinity of the gene encoding the A-type inclusion body protein. The possible functions of the analyzed viral proteins are discussed.

Published
1994

8. [Study of the structure-activity organization of the variola virus genome. II. Analysis of the nucleotide sequence of the HindIII region (C,E,R,Q,K and H)-DNA fragments of the India-1967 strain].

Author

Shchelkunov SN, Blinov VM, Resenchuk SM, Gutorov VV, Safronov PF, Kurmanov RK, Totmenin AV, Chizhikov VE, Marennikova SS, and Sandakhchiev LS

Subjects
Amino Acid Sequence, Molecular Sequence Data, Restriction Mapping, Sequence Homology, Amino Acid, Structure-Activity Relationship, DNA, Viral genetics, Genome, Viral, Variola virus genetics
Abstract

Sequencing of variola virus (VAR) genome region of 43069 bp was carried out. This area contains 42 potential genes. Computer analysis of proteins coding for these viral genes was done. We compared VAR proteins with the those of vaccinia virus. The region studied is conservative for orthopoxviruses.

Published
1993

9. [Study of the structural-functional organization of the natural variola virus genome. I. Cloning HindIII- and XhoI-fragments of viral DNA and sequencing HindIII -M, -L, -I fragments].

Author

Shchelkunov SN, Blinov VM, Totmenin AV, Marennikova SS, Kolykhalov AA, Frolov IV, Chizhikov VE, Gutorov VV, Gashnikov PV, and Belanov EF

Subjects
Amino Acid Sequence, Base Sequence, Cloning, Molecular, Cosmids, Deoxyribonuclease HindIII genetics, Deoxyribonucleases, Type II Site-Specific genetics, Genes, Viral, Molecular Sequence Data, Plasmids, Restriction Mapping, Structure-Activity Relationship, DNA, Viral genetics, Variola virus genetics
Abstract

HindIII and XhoI genome fragments of variola major virus strain India-1967 were inserted into the bacterial plasmids and cosmid. Sequencing and computer analysis of the region of HindIII M, L, and I DNA fragments of the virus studied have been carried out.

Published
1992

10. [Molecular-biological study of vaccinia virus genome. I. Cloning of vaccinia virus DNA fragments in bacterial vectors].

Author

Riazankina OI, Totmenin AV, Shchelkunov SN, and Malygin EG

Subjects
Blotting, Southern, DNA, Viral chemistry, Deoxyribonuclease HindIII, Escherichia coli genetics, Genetic Vectors, Plasmids, Restriction Mapping, Cloning, Molecular, DNA, Viral genetics, Genes, Viral, Vaccinia virus genetics
Abstract

The HindIII DNA fragments of vaccinia virus strain L-IVP were cloned in pBR322 bacterial plasmid. A hybrid plasmids collection of pVHn series contains all fragments of virus genome except terminal HindIII-B and HindIII-G, and also a large HindIII-A. The latter was cloned in cosmid pHC79. The obtained collection of hybrid DNA molecules allows to carry out a wide range of molecular biological experiments on the vaccinia virus genome.

Published
1990

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