RESUMO
The recombinant plasmids containing the gene for hepatitis B viral core-antigen with the pre-core-sequence controlled by the early-late promoter of the 7.5' K protein gene were constructed. The recombinant strains of vaccinia virus were obtained on their basis (vHBe42-1 and vHBe42-3) selectively expressing HBeAg of hepatitis B virus. The kinetics of HBeAg synthesis was studied in infected cells as well as secretion of the protein into culturing medium. Three proteins were found by blotting technique in the cells infected by vHBe42-3 that react with the specific antiserum to HBeAg and have the mol. masses 25, 22 and 17 kD. The completely processed HBeAg 17 kD was found in the culturing medium. The rabbit serums from the animals immunized by recombinant vHBe42-3 contained antibodies to HBeAg but not to HBcAg. This makes it possible to study the structural and functional organization, immunological properties and role of this antigen in pathogenesis of hepatitis virus B and to construct the specific test systems for screening HBeAg and corresponding antibodies.
Assuntos
Antígenos E da Hepatite B/biossíntese , Recombinação Genética , Vaccinia virus/genética , Animais , Western Blotting , Bovinos , Células Cultivadas , DNA Viral/genética , Escherichia coli/enzimologia , Expressão Gênica , Genes Virais , Cinética , Plasmídeos , Mapeamento por Restrição , Transfecção , Proteínas Virais/análise , beta-Galactosidase/análiseAssuntos
Capsídeo/biossíntese , Antígenos do Núcleo do Vírus da Hepatite B/biossíntese , Vaccinia virus/metabolismo , Capsídeo/genética , Capsídeo/imunologia , DNA Recombinante , Epitopos/biossíntese , Epitopos/genética , Antígenos do Núcleo do Vírus da Hepatite B/genética , Antígenos do Núcleo do Vírus da Hepatite B/imunologia , Vacinas contra Hepatite B , Immunoblotting , Plasmídeos , Vacinas Sintéticas/biossíntese , Vacinas Sintéticas/genética , Vaccinia virus/genética , Vacinas contra Hepatite Viral/biossíntese , Vacinas contra Hepatite Viral/genética , Vacinas contra Hepatite Viral/imunologiaRESUMO
Ten recombinants between the viruses of vaccinia and ectromelia were isolated that cause the ectromelia virus specific lesions in mice. The structure of recombinant viral genomes, the efficiency of viral propagation in mice, the nature of lesions induced by viruses have been studied. Eight of obtained recombinants have a DNA insertion originating from the right end of ectromelia viral genome, nine recombinants have an insertion originating from the left end, seven recombinants possess both insertions. The latter recombinants have more pronounced pathogenicity for mice. Both revealed regions are supposed to define the specific pathogenicity of ectromelia virus for mice.
