Development of a method for markerless gene deletion in Pseudomonas putida.

  title={Development of a method for markerless gene deletion in Pseudomonas putida.},
  author={Nadja Graf and Josef Altenbuchner},
  journal={Applied and environmental microbiology},
  volume={77 15},
We developed a negative counterselection system for Pseudomonas putida based on uracil phosphoribosyltransferase (UPRTase) and sensitivity against the antimetabolite 5-fluorouracil (5-FU). We constructed a P. putida strain that is resistant to 5-FU and constructed vectors for the deletion of the surface adhesion protein gene, the flagellum biosynthesis operon, and two endonuclease genes. The genes were efficiently disrupted and left a markerless chromosomal in-frame deletion. 
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