Submitted to: Molecular Genetics of Bacteria and Phage
Publication Type: Abstract Only
Publication Acceptance Date: 6/8/2011
Publication Date: 8/2/2011
Citation: Swingle, B.M. 2011. New strategies for genetic engineering Pseudomonas syringae using recombination. Molecular Genetics of Bacteria and Phage. p. 20.
Technical Abstract: Here we report that DNA oligonucleotides (oligos) introduced directly into bacteria by electroporation can recombine with the bacterial chromosome. This phenomenon was identified in Pseudomonas syringae and we subsequently found that Escherichia coli, Salmonella typhimurium and Shigella flexneri are also capable of oligo recombination. Parameters such as oligo concentration and length were evaluated in terms of the influence on recombination frequency. This phenomenon has many potential uses. Here we describe two applications of this technology including a method to characterize the genetic basis for antibiotic resistance. This was demonstrated by examining the rpoB mutation responsible for rifampicin resistance in P. syringae pv. tomato DC3000. Secondly, we describe the use of this system to identify bacterial enzymes that catalyze elevated levels of recombination reactions.