Data from: A platform supporting generation and isolation of random transposon mutants in Chlamydia trachomatis

Chlamydia species represent a paradigm for understanding successful
obligate intracellular parasitism. Despite limited genetic malleability,
development of genetic tools has facilitated the elucidation of molecular
mechanisms governing infectivity. Random mutagenesis approaches provide
one of the most powerful strategies available to accomplish untargeted
elucidation of gene function. Unfortunately, initial progress in
transposon-mediated mutagenesis of Chlamydia has been challenging. To
increase efficiency, we developed a plasmid-based system that couples
conditional plasmid maintenance with a previously described strategy
leveraging inducible expression of the Himar1-derived C9 transposase. Our
pOri-Tn(Q) construct was maintained in C. trachomatis cultivated with
antibiotics but was rapidly cured in the absence of antibiotic selection.
pOri-Tn(Q) supported transposition events when transposase expression was
induced during infection. Induction was accompanied by loss of the plasmid
backbone when Penicillin G was used to select for only the transposable
element. C9 induction during iterative passaging was used to increase
overall insertion frequency and accumulate an expanded pool of transposon
mutants. The approach supported isolation of individual mutant strains
from the mixed pool, and whole-genome sequencing confirmed that the
recovered strains harbored single insertions.