title: Site directed chromosomal marking of a fluorescent pseudomonad isolated from the phytosphere of sugar beet; stability and potential for marker gene transfer.
abstract: A plasmid-free, non-pathogenic, ribosomal RNA group 1 fluorescent pseudomonad, Pseudomonas fluorescens SBW25, was selected from the microflora of sugar beet (Beta vulgaris) and modified to contain constitutively expressed marker genes. By site directed homologous recombination a KX cassette [kanamycin resistance (kanr) and catechol 2,3 dioxygenase (xylE)] and a ZY cassette [lactose utilization (lacZY, beta-galactosidase, lactose permease)] were introduced at least 1 Mbp apart on the 6.6 Mbp bacterial chromosome. Separate sites were selected to provide sensitive detection methods and allow assessments of marker gene stability of the genetically modified micro-organism (GMM), SBW25EeZY6KX, when it colonized the leaves and roots of sugar beet plants following seed inoculation.
authors.affiliation: Natural Environment Research Council, Institute of Virology and Environmental Microbiology, Oxford, UK.
authors.forename: M J
authors.initials: MJ
authors.lastname: Bailey
authors.forename: A K
authors.initials: AK
authors.lastname: Lilley
authors.forename: I P
authors.initials: IP
authors.lastname: Thompson
authors.forename: P B
authors.initials: PB
authors.lastname: Rainey
authors.forename: R J
authors.initials: RJ
authors.lastname: Ellis
mesh_terms: D001483:Base Sequence; D002874:Chromosome Mapping; D002876:Chromosomes, Bacterial / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D017753:Ecosystem; D018014:Gene Transfer Techniques; D005819:Genetic Markers; D007763:Lac Operon; D008969:Molecular Sequence Data; D011551:Pseudomonas fluorescens / Q000235:genetics* / Q000302:isolation & purification*; D011995:Recombination, Genetic; D014675:Vegetables / Q000382:microbiology*
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D005819:Genetic Markers