Development of a Method for Markerless Gene Deletion in Pseudomonas putida
- 1 August 2011
- journal article
- Published by American Society for Microbiology in Applied and Environmental Microbiology
- Vol. 77 (15) , 5549-5552
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.Keywords
This publication has 15 references indexed in Scilit:
- Development of a Markerless Genetic Exchange System for Desulfovibrio vulgaris Hildenborough and Its Use in Generating a Strain with Increased Transformation EfficiencyApplied and Environmental Microbiology, 2009
- Development and Application of a upp -Based Counterselective Gene Replacement System for the Study of the S-Layer Protein SlpX of Lactobacillus acidophilus NCFMApplied and Environmental Microbiology, 2009
- Functional Genomics of Stress Response in Pseudomonas putida KT2440Journal of Bacteriology, 2006
- Development of a Method for Markerless Genetic Exchange in Enterococcus faecalis and Its Use in Construction of a srtA MutantApplied and Environmental Microbiology, 2005
- Growing and Analyzing Static BiofilmsCurrent Protocols in Microbiology, 2005
- Complete genome sequence and comparative analysis of the metabolically versatile Pseudomonas putida KT2440Environmental Microbiology, 2002
- Broad-Host-Range cre-lox System for Antibiotic Marker Recycling in Gram-Negative BacteriaBioTechniques, 2002
- An improved system for gene replacement and xylE fusion analysis in Pseudomonas aeruginosaGene, 1995
- [40] Positive selection vectors based on palindromic DNA sequences1992
- Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectorsGene, 1985