ABSTRACT
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.
Subject(s)
Antigens, Bacterial/analysis , Bordetella pertussis/immunology , Antigens, Bacterial/chemistry , Antigens, Bacterial/isolation & purification , Bordetella pertussis/growth & development , Chemical Phenomena , Chemistry, Physical , Culture Media , Electrophoresis, Polyacrylamide Gel , Hydrogen-Ion Concentration , Molecular Weight , Peptides/analysis , Time FactorsABSTRACT
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.
Subject(s)
Bacillus/enzymology , DNA Restriction Enzymes/isolation & purification , Deoxyribonucleases, Type II Site-Specific , Base Sequence , Chromatography, Ion Exchange , DNA Restriction Enzymes/genetics , DNA Restriction Enzymes/metabolism , Electrophoresis, Cellulose Acetate , Substrate SpecificityABSTRACT
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.
Subject(s)
DNA Restriction Enzymes/isolation & purification , Deoxyribonucleases, Type II Site-Specific , Proteus mirabilis/enzymology , Bacteriophage lambda , DNA, ViralABSTRACT
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.
Subject(s)
Bordetella pertussis/genetics , Cloning, Molecular , DNA, Bacterial/genetics , Escherichia coli/genetics , Base Sequence , Gene Expression Regulation , Genes, Bacterial , Genetic Vectors , Nucleic Acid Hybridization , Plasmids , Protein BiosynthesisABSTRACT
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.
