I am PhD scholar, interesting to work on resistance mechanism of PDR K. pneumonia, particularly on plasmid mediated. I have tried ethidium bromide and SDS for plasmid curing phenotypically, but no result was obtained.
If you know anything about the sequence of the plasmid I imagine you could try knocking out important parts for replication (like a rep protein, if your plasmid has one) with λred. That way you also would have less chance of secondary mutations to the chromosome. There's lambda recombination vectors that have selection markers like Hygromycin B and Blasticidin resistance, antibiotics which are both toxic to eukaryotes and aren't used clinically so hopefully your strain won't be resistant to them:
https://redrecombineering.ncifcrf.gov/strains--plasmids.html
You would definitely have to play around with MIC concentrations for your strain though, sometimes the working concentration for cloning strains of E. coli is a lot less than what it is for wild-type strains of other species (especially highly resistant ones.)
Hello Muhammad, SDS-based curing method is a very effective method because plasmid-bearing cells are highly sensitive due to the presence of plasmid-specified pili on their surfaces. https://scinapse.io/papers/2036708782
However, specific plasmid curing experiment on Klebsiela pneumoniae could be done by exposing the cells to sub-minimum inhibitory concentration (sub-MIC) of acridine orange. You could also try successive bacterial subculturing at 44 degC.
Effective and cheap method for expression of efflux pump genes in Gram negative MDRs.
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