RESUMO
A virus, identified as cowpox virus by its biological properties and the results of the analysis of its DNA, was isolated from a sick 4-year-old child with a clinical picture of pox, though having had no contacts with known natural carriers of the causative agent of this infection. At the same time the isolated virus was found to differ from the reference strain, as well as from other isolates of vaccinia virus by some biological markers (and in particular by the structure of cytoplasmic inclusions of type A) and by the restriction profile of DNA. The Hind III maps indicating the location of restriction sites made it possible to localize the genome differences established in this study. The specific feature of this case was the previous close contact of the child with a mole which was probably the source of infection.
Assuntos
Vírus da Varíola Bovina/classificação , Varíola Bovina/virologia , Animais , Embrião de Galinha , Pré-Escolar , Chlorocebus aethiops , Varíola Bovina/patologia , Vírus da Varíola Bovina/genética , Vírus da Varíola Bovina/isolamento & purificação , Vírus da Varíola Bovina/patogenicidade , DNA Viral/genética , Feminino , Humanos , Camundongos , Microscopia Eletrônica , Ratos , Mapeamento por Restrição/métodos , Células VeroRESUMO
Two genetic markers--the thymidine kinase gene of herpes simplex virus, and the beta-galactosidase gene of Escherichia coli--were incorporated into the 36K protein gene (IL1 gene according to the nomenclature of the Copenhagen strain of vaccinia virus; Goebel et al., 1990) from the HindIII-P DNA fragment of the LIVP strain (variant of Lister strain) of vaccinia virus (VV). After recombination of the obtained integration plasmid pVZ64-TK with the VV genome (tk-), it was found that the resultant TK+ viruses were unstable with respect to the Lac+ phenotype. On the basis of hybridization of DNA fragments of selected clones, a scheme for the formation of hybrid viruses is proposed, and an approach to a simple phenotypical discrimination between essential and non-essential genes for VV viability is described.
Assuntos
Vaccinia virus/fisiologia , Proteínas não Estruturais Virais/genética , Proteínas Virais/genética , Replicação Viral/genética , Animais , Células Cultivadas , Chlorocebus aethiops , DNA Viral , Genoma Viral , Fenótipo , Plasmídeos , Ratos , Recombinação Genética , Vaccinia virus/genética , Integração ViralRESUMO
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.
Assuntos
DNA Viral/genética , Vírus da Varíola/genética , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , Cosmídeos , Desoxirribonuclease HindIII/genética , Desoxirribonucleases de Sítio Específico do Tipo II/genética , Genes Virais , Dados de Sequência Molecular , Plasmídeos , Mapeamento por Restrição , Relação Estrutura-AtividadeRESUMO
A hybrid vaccinia virus expressing a chimeric protein consisting of thymidine kinase and the encephalitogenic determinant, S1, from guinea pig myelin basic protein was constructed. Infection of guinea pigs with the virus resulted in the development of allergic encephalomyelitis.
