Development of a Method for Markerless Gene Deletion in Pseudomonas putida | Zendy

Nadja Graf | Zendy; Josef Altenbuchner | Zendy

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Development of a Method for Markerless Gene Deletion in Pseudomonas putida

Author(s) -

Nadja Graf,

Josef Altenbuchner

Publication year - 2011

Publication title -

applied and environmental microbiology

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 1.552

H-Index - 324

eISSN - 1070-6291

pISSN - 0099-2240

DOI - 10.1128/aem.05055-11

Subject(s) - pseudomonas putida , biology , operon , gene , genetics , recombineering , uracil , plasmid , dna , escherichia coli

We developed a negative counterselection system forPseudomonas putida based on uracil phosphoribosyltransferase (UPRTase) and sensitivity against the antimetabolite 5-fluorouracil (5-FU). We constructed aP. 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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