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Characterization of Two Inducible Bacteriophages, α 1 and α 2, Isolated from Clostridium botulinum Type A 190L and Their Deoxyribonucleic Acids
Author(s) -
Kinouchi Takemi,
Takumi Kenji,
Kawata Tomio
Publication year - 1981
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.1981.tb00096.x
Subject(s) - biology , dna , ecori , restriction enzyme , lysogenic cycle , microbiology and biotechnology , clostridium botulinum , bacteriophage , strain (injury) , plasmid , restriction fragment , biochemistry , gene , escherichia coli , toxin , anatomy
Two inducible bacteriophages, α 1 and α 2, isolated from Clostridium botulinum type A strain 190L and their deoxyribonucleic acids (DNAs) were purified and characterized. Phage α l, which is unable to form plaques on any strain of C. botulinum , was produced in large quantities after treatment with mitomycin C (MC), whereas phage α 2, which was induced in much lower quantities than phage α l, propagated in cultures of type A strain Hall. The phage DNAs were exclusively synthesized after induction with MC. α l and α 2 DNAs had sedimentation coefficients of 34.0 and 30.6 S, corresponding to molecular weights of 31.9 × 10 6 and 23.5 × 10 6 , respectively. The buoyant density in CsCl was 1.682 g/cm 3 for al DNA and 1.680 g/cm 3 for α 2 DNA. Based on thermal denaturation characteristics, the genomes of both phages were shown to be double‐stranded DNAs. Agarose gel electrophoretic profiles of the phage DNAs digested with restriction endonuclease EcoRI revealed nine fragments for al DNA and six fragments for α 2 DNA. The molecular weights of the phage DNAs as determined by restriction enzyme analysis were 30.55 × 10 6 for α 1 DNA and 25.83 × 10 6 for α 2 DNA. Nontoxigenic mutants obtained from strain 190L could, like the toxigenic parent strain, produce the two phages after treatment with MC. Lysogenic conversion to toxigenicity by phage α 2 was not observed with the nontoxigenic mutants. It seems likely that there is no relationship between either phage genome and the toxigenicity of C. botulinum type A.