Transposon insertion and subsequent donor formation promoted by Tn501 in Bordetella pertussis

Abstract

The mercuric chloride resistance transposon, Tn501, was introduced into Bordetella pertussis by using the chimeric plasmid pUW942, which is unable to replicate in this species. Tn501 insertions which conferred a thiamine requirement were the predominant insertion class. In many cases, the mercuric chloride-resistant transconjugants were also resistant to the other plasmid markers, but failure to detect plasmid DNA in these isolates indicated that integration of the entire plasmid into the chromosome had occurred. One such insertion was further characterized. Southern hybridization with a Tn501-specific probe indicated that chromosomal DNA from one strain containing the integrated plasmid had two copies of Tn501 and an intervening copy of the plasmid associated with the chromosome. The presence of the plasmid was unstable, and derivatives which had lost all of the plasmid markers except mercuric chloride resistance were obtained. These strains had a single copy of Tn501 and had lost all of the rest of the plasmid-specific sequences. Strains containing the plasmid in the integrated state could act as genetic donors and mobilize chromosomal genes.