Genetic Aspects of Clostridium Botulinum
The relationship of bacteriophages and plasmids to the production of neurotoxins was studied in strains of Clostridium botulinum types A through G. Neurotoxins C1 and D produced by types C and D, respectively, were shown to be mediated by specific bacteriophages. Evidence is presented that strongly suggests that both neurotoxin and bacteriocin production by type G are in some manner related to a 81-MDa plasmid carried by toxigenic strains.
Antigenicity and host range of four type C and three type D converting phages were studied. The phages were classified into three groups based on their antigenicity and host range: group 1 consisted of c-st and c-468 phages; group 2 was c-203, c-d6f, and d-1873: and group 3 was d-sa and d-4947.
Nucleic acids were extracted from groups 1 and 2 phages, and noncon- verting mutant phage (c)-n71 which was obtained from C-Stockholm strain as well as c-st phage. The susceptibility of phage DNAs to different types of nucleases was observed. It was concluded that the nucleic acids of all six phages were double-stranded DNA. The length of c-st, (c)-n71, c-468, and c-d6f phage DNAs was about 110 kilobase pairs and that of c-203 and d-1873 was 150 kilobase pairs. PstI digested the DNAs from two group 1 phages and (c)-n71 phage with very similar patterns, but did not digest the DNAs from group 2 phages. On the contrary, Sau3A digested only the DNAs from group 2 phages though the similarity of digestion patterns was low.
The existence of the structure genes for the toxin in these five converting phages belonging to groups 1 and 2 and (c)n-71 was confirmed by the hybridization the DNA sequence predicted for the N-terminal amino acids (2 to 17) of C. botulinum type C toxin. The loss of the converting ability of (c)-n71 phage may be caused not by the delection of tox+ gene but rather by the base mutation in c-st phage DNA.
KeywordsPhenol Agar Agarose Chloroform Polyethylene
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