RESUMO
Some physicochemical properties of B.pertussis antigenic complexes isolated from synthetic and semisynthetic culture media have been studied. The chemical composition of these complexes has been determined. Proteins, polypeptide subunits and lipopolysaccharide forming these complexes have been characterized. The presence of two main protective substances of B.pertussis has been revealed: fimbrial hemagglutinin and B.pertussis toxin. The influence of culture medium on the level of the synthesis of B.pertussis adenylate cyclase has been studied.
Assuntos
Antígenos de Bactérias/análise , Bordetella pertussis/imunologia , Antígenos de Bactérias/química , Antígenos de Bactérias/isolamento & purificação , Bordetella pertussis/crescimento & desenvolvimento , Fenômenos Químicos , Físico-Química , Meios de Cultura , Eletroforese em Gel de Poliacrilamida , Concentração de Íons de Hidrogênio , Peso Molecular , Peptídeos/análise , Fatores de TempoRESUMO
A strain producing the new specific restriction endonuclease BcmI has been found in the Bacillus generum. The enzyme has been purified by chromatography on the blue sepharose, phosphocellulose PII, heparin sepharose. The analogous purification has been obtained when the blue sepharose has been substituted for the orange sepharose, the home produced sorbent. The BcmI enzyme has been shown by the substrate specificity definition to be an isoschizomer of the restriction endonuclease ClaI.
Assuntos
Bacillus/enzimologia , Enzimas de Restrição do DNA/isolamento & purificação , Desoxirribonucleases de Sítio Específico do Tipo II , Sequência de Bases , Cromatografia por Troca Iônica , Enzimas de Restrição do DNA/genética , Enzimas de Restrição do DNA/metabolismo , Eletroforese em Acetato de Celulose , Especificidade por SubstratoRESUMO
A new restriction endonuclease Pmi I was detected in Proteus mirabilis 1667. The enzyme hydrolyzes DNA of the phage lambda into 10 electrophoretically separating fragments with molecular weights of 1.3-7.9 mD. With the use of two-stage chromatography on blue sepharose and phosphocellulose it is possible to obtain restriction endonuclease Pmi I free of the admixtures of ballast proteins, nonspecific nucleases and phosphatases.
Assuntos
Enzimas de Restrição do DNA/isolamento & purificação , Desoxirribonucleases de Sítio Específico do Tipo II , Proteus mirabilis/enzimologia , Bacteriófago lambda , DNA ViralRESUMO
Hybrid plasmids containing B. pertussis DNA insertions have been constructed with the use of the vector plasmid pBR 322 and the Pst I fragments of B. pertussis DNA. Some properties of the hybrid plasmids are characterized. The possibility of the expression of B. pertussis genes in the protein-synthetizing cell-free system obtained from E. coli has been demonstrated.
Assuntos
Bordetella pertussis/genética , Clonagem Molecular , DNA Bacteriano/genética , Escherichia coli/genética , Sequência de Bases , Regulação da Expressão Gênica , Genes Bacterianos , Vetores Genéticos , Hibridização de Ácido Nucleico , Plasmídeos , Biossíntese de ProteínasRESUMO
New restriction endonuclease has been isolated from Bordetella pertussis vaccine strain 305 and purified in 1 stage on Sepharose covalently bound with blue dextran. The isolated restrictase has been found capable of breaking down lambda-phag DNA into 7 fragments. According to its specificity, Bpe I is the isoschizomer of Hind III obtained from Haemophilus influenzae strain Rd.
