Site directed chromosomal marking of a fluorescent pseudomonad isolated from the phytosphere of sugar beet; stability and potential for marker gene transfer.
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.
Bailey
M J
MJ
Natural Environment Research Council, Institute of Virology and Environmental Microbiology, Oxford, UK.
Lilley
A K
AK
Thompson
I P
IP
Rainey
P B
PB
Ellis
R J
RJ
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*
D017931:DNA Primers; D004269:DNA, Bacterial; D005819:Genetic Markers