title: Development of a method for markerless gene deletion in Pseudomonas putida.
abstract: 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.
authors.affiliation: Institut für Industrielle Genetik, Universität Stuttgart, Allmandring 31, 70569 Stuttgart, Germany.
authors.forename: Nadja
authors.initials: N
authors.lastname: Graf
authors.forename: Josef
authors.initials: J
authors.lastname: Altenbuchner
mesh_terms: D001422:Bacterial Adhesion / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D024881:Drug Resistance, Bacterial; D004720:Endonucleases / Q000235:genetics; D005407:Flagella / Q000235:genetics; D005472:Fluorouracil / Q000494:pharmacology*; D005821:Genetic Techniques; D010430:Pentosyltransferases / Q000235:genetics*; D010957:Plasmids; D016958:Pseudomonas putida / Q000235:genetics* / Q000378:metabolism; D017384:Sequence Deletion
chemical_list: D001426:Bacterial Proteins; D010430:Pentosyltransferases; C023269:uracil phosphoribosyltransferase; D004720:Endonucleases; D005472:Fluorouracil