Isolation, characterization, and utilization of a temperature-sensitive allele of a Pseudomonas replicon.

@article{Silosuh2009IsolationCA,
  title={Isolation, characterization, and utilization of a temperature-sensitive allele of a Pseudomonas replicon.},
  author={Laura Silo-suh and Brett Elmore and Dennis E. Ohman and Sang-Jin Suh},
  journal={Journal of microbiological methods},
  year={2009},
  volume={78 3},
  pages={319-24}
}
In order to facilitate genetic study of the opportunistic bacterial pathogen Pseudomonas aeruginosa, we isolated a conditional, temperature-sensitive plasmid origin of replication. We mutagenized the popular Pseudomonas stabilizing fragment from pRO1610 in vitro using the Taq thermostable DNA polymerase in a polymerase chain reaction (PCR). Out of approximately 23,000 potential clones, 48 temperature-sensitive mutants were isolated. One mutant was further characterized and the origin of… CONTINUE READING