RESUMO
Libraries of hybrid plasmids carrying DNA fragments of complete genomes of 8 variola virus strain from the Russian Collection belonging to 2 epidemical types and isolated in various geographic regions of the world were obtained. Genomic sequences of variola virus can be thus preserved for a long time in a biologically safe form and provide the research work on studying the genetic organization of this unique virus and on developing modern methods for rapid detection of variola virus and other orthopoxviruses.
Assuntos
Genoma Viral , Vírus da Varíola/genética , DNA Viral/análise , DNA Viral/genética , Saúde Global , Plasmídeos/genética , Reação em Cadeia da Polimerase , Mapeamento por RestriçãoRESUMO
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.
Assuntos
Genoma Viral , Monkeypox virus/genética , Monkeypox virus/patogenicidade , Vírus da Varíola/genética , Vírus da Varíola/patogenicidade , Sequência de Aminoácidos , Anquirinas/química , Evolução Molecular , Humanos , Modelos Genéticos , Dados de Sequência Molecular , Fases de Leitura Aberta , Filogenia , Estrutura Terciária de Proteína , Homologia de Sequência de Aminoácidos , VirulênciaRESUMO
Poxviruses, belonging to the genus of Orthopoxviruses and Leporipoxviruses, express a protein that binds a wide spectrum of CC-chenokines. The above viral protein (vCCI) has no homology by the amino acid sequence with the known cellular receptors of chemokines or with any other proteins represented in databases. Sixty-nine nucleotide sequences of the vCCI protein gene of 4 Orthopoxviruses, which are pathogenic to man, and of 2 species, to whom man is immune, as well as 16 of such sequences borrowed from GenBank were subjected to the phylogenetic analysis. Strains of Orthopoxviruses, except for cowpox viruses (CPXV), were found to cluster together in groups according to their specific attribute. CPXV isolates can be conditionally divided into 2 subgroups, which are much remote from one another evolutionally. Four subgroups were defined for smallpox virus with their strains originating from different geographic regions.
Assuntos
Quimiocinas/metabolismo , Orthopoxvirus/genética , Proteínas Virais/genética , Sequência de Bases , DNA Viral , Dados de Sequência Molecular , Filogenia , Ligação Proteica , Homologia de Sequência do Ácido NucleicoRESUMO
A method for describing the Orthopoxviruses that are pathogenic both to man and animals is described in the article. The method is based on hybridization of a fluorescently labelled amplified DNA sample with oligonucleotides, which were immobilized in a microchip. Species-specific regions within the crmB gene encoding a viral analogue of the tumor necrosis factor receptor, i.e. an important gene determining the pathogenicity of the mentioned Orthopoxviruses type, were used as a target for identification. The identification procedure takes around 6 hours and does not demand any costly equipment (a portable fluorescent microscope can be used).
Assuntos
Análise de Sequência com Séries de Oligonucleotídeos/métodos , Orthopoxvirus/isolamento & purificação , Receptores do Fator de Necrose Tumoral/genética , Proteínas Virais/genética , Animais , Sequência de Bases , DNA Viral/análise , Perfilação da Expressão Gênica , Humanos , Dados de Sequência Molecular , Sondas de Oligonucleotídeos , Orthopoxvirus/genética , Orthopoxvirus/patogenicidade , Alinhamento de Sequência , Especificidade da EspécieRESUMO
Monkeypox virus (MPV) belongs to the orthopoxvirus genus of the family Poxviridae, is endemic in parts of Africa, and causes a human disease that resembles smallpox. The 196,858-bp MPV genome was analyzed with regard to structural features and open reading frames. Each end of the genome contains an identical but oppositely oriented 6379-bp terminal inverted repetition, which similar to that of other orthopoxviruses, includes a putative telomere resolution sequence and short tandem repeats. Computer-assisted analysis was used to identify 190 open reading frames containing >/=60 amino acid residues. Of these, four were present within the inverted terminal repetition. MPV contained the known essential orthopoxvirus genes but only a subset of the putative immunomodulatory and host range genes. Sequence comparisons confirmed the assignment of MPV as a distinct species of orthopoxvirus that is not a direct ancestor or a direct descendent of variola virus, the causative agent of smallpox.