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Advanced approaches to the synthesis and reconstruction of genetic material developed in the Institutes of Molecular Biology and Genetics during the past years are summarized. The evolution of methods for oligonucleotide synthesis and scopes for their use in gene production are discussed. The principles of localised mutagenesis methods developed in the Institute are described, such as: a) mutagenesis directed to the regulatory gene regions; b) segment-localized mutagenesis; c) mutagenesis directed by phosphotriester analogues of oligonucleotides. Examples of employing these methods for induction of regulatory mutants of phage lambda, production of fused genes, mutant interferon genes, construction of new DNA vectors, construction of hybrid H1-H3 subtype haemagglutinine gene of influenza virus etc. are presented. The approach to in vivo site-directed mutagenesis is experimentally substantiated. 相似文献
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Using immunochemical and, subsequently, autoradiographic methods, the authors have shown that after the injection of the total fraction of polyribosomes isolated from the rat liver in the Acetabularia cytoplasm a protein interacting with the antiserum against rat thyrosine aminotransferase is synthesized. Prospects of using Acetabularia as a test-system for the translation of heterogenous mRNA and mRNP are discussed. 相似文献
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Tumanova OIu Kuvshinov VN Azaev MSh Masharskiĭ AE Klimov NA Kozlov AP Il'ichev AA Sandakhchiev LS 《Molekuliarnaia biologiia》2001,35(1):146-151
A phase peptide library was screened with virus-neutralizing monoclonal antibodies (MCA) 2F5 which recognize a conserved epitope of HIV-1 gp41. Phages that expose peptides specifically binding with MCA 2F5 were selected by ELISA. Amino acid sequence analysis revealed a homology to region 662-671 of HIV-1 HB10 gp160 for most peptides. The major role in recognition was ascribed to Asp-664, Lys-665, and Trp-666. The epitope-mimicking peptides were tested for immunogenicity. Antibodies to gp41 were detected in serum of immunized rabbits. 相似文献
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Tat'kov SI Smirnova OYu Tsivkovskii RYu Kochneva GV Kuz'micheva GA Khristoforov VS Kosarev IS Chernykh ER Khonina NA Lebedev LR Danilenko ED Fadina VA Pustoshilova NM Masycheva VI Sandakhchiev LS 《Doklady biochemistry : proceedings of the Academy of Sciences of the USSR, Biochemistry section / translated from Russian》2000,372(1-6):112-114
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Human monkeypox and smallpox viruses: genomic comparison. 总被引:8,自引:0,他引:8
S N Shchelkunov A V Totmenin I V Babkin P F Safronov O I Ryazankina N A Petrov V V Gutorov E A Uvarova M V Mikheev J R Sisler J J Esposito P B Jahrling B Moss L S Sandakhchiev 《FEBS letters》2001,509(1):66-70
Monkeypox virus (MPV) causes a human disease which resembles smallpox but with a lower person-to-person transmission rate. To determine the genetic relationship between the orthopoxviruses causing these two diseases, we sequenced the 197-kb genome of MPV isolated from a patient during a large human monkeypox outbreak in Zaire in 1996. The nucleotide sequence within the central region of the MPV genome, which encodes essential enzymes and structural proteins, was 96.3% identical with that of variola (smallpox) virus (VAR). In contrast, there were considerable differences between MPV and VAR in the regions encoding virulence and host-range factors near the ends of the genome. Our data indicate that MPV is not the direct ancestor of VAR and is unlikely to naturally acquire all properties of VAR. 相似文献
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Immunogenic Properties of Peptides Mimicking a HIV-1 gp41 Epitope Recognized by Virus-Neutralizing Antibodies 2F5 总被引:1,自引:0,他引:1
Tumanova O. Yu. Kuvshinov V. N. Orlovskaya I. A. Pronyaeva T. R. Pokrovskii A. G. Il'ichev A. A. Sandakhchiev L. S. 《Molecular Biology》2003,37(3):473-476
Phage display was used to obtain peptides mimicking a HIV-1 gp41 conserved epitope recognized by virus-neutralizing monoclonal antibodies (mAb) 2F5. Rabbits and mice were immunized with the peptides exposed on the surface of filamentous bacteriophages. Antibodies to gp41 were detected in the sera of immunized animals. The virus-neutralizing activity of the sera was examined. 相似文献