Open AccessMitomycin C induction of bacteriophages from Serpulina hyodysenteriae and Serpulina innocens
Author(s) -
Humphrey Sam B.,
Stanton Thad B.,
Jensen Neil S.
Publication year - 1995
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1995.tb07921.x
Subject(s) - mitomycin c , microbiology and biotechnology , biology , prophage , lysis , bacteriophage , bacteria , lysogenic cycle , dna , agarose gel electrophoresis , gel electrophoresis , agarose , escherichia coli , genetics , gene
A prophage was induced from cells of the pathogenic spirochaete Serpulina hyodysenteriae using mitomycin C. Five to seven hours after mitomycin C was added (8 μg/ml, final concentration) to S. hyodysenteriae B204 cultures in BHIS broth (OD 620 = 0.9) cell lysis was detected as a decrease in culture optical density. Bacteriophage particles attached to whole cells and to cell debris were detected by electron microscopic analysis of negatively stained (2% PTA, pH 7.0) bacteria harvested by centrifugation from mitomycin C treated cultures. The phage particles consisted of a head (45 nm diameter) and a tail (64 nm × 9 nm). Bacteria from untreated cultures lacked phages detectable by electron microscopy. The appearance of bacteriophage particles in mitomycin C treated cultures correlated with the appearance of extrachromosomal DNA, 7–8 kb in size as estimated by agarose gel electrophoresis, in DNA preparations from treated S. hyodysenteriae cells. When cultures of other S. hyodysenteriae strains (B78, B169, A‐1, B8044, B6933, Ack300/8, R‐1) and S. innocens 4/71 in BHIS were treated with mitomycin C (8–15 μg/ml, final concentration), phages similar in morphology and size to the S. hyodysenteriae B204 phage were induced.