Tracking Recombinant Organisms in the Environment: β–Galactosidase as a Selectable Non–Antibiotic Marker for Fluorescent Pseudomonads

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Abstract

A sensitive, selectable marker system has been developed for fluorescent pseudomonads based on expression of the E. coli lac operon genes encoding β–galactosidase and lactose permease. Examination of over 500 fluorescent Pseudomonas isolates from soil samples revealed the constant absence of the o–nitrophenylgalactoside (ONPG) positive phenotype, and the inability to utilize lactose as a sole carbon source. Based on these observations broad host–range plasmids containing E. coli lacZ and lacY genes were constructed and stably introduced into native P. fluorescens strains to confer the ability to cleave the chromogenic substrate X–Gal and to grow on minimal lactose media. This marker system enabled the detection of Lac+ transformants at a sensitivity of <10 CFU/g soil.

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Drahos, D., Hemming, B. & McPherson, S. Tracking Recombinant Organisms in the Environment: β–Galactosidase as a Selectable Non–Antibiotic Marker for Fluorescent Pseudomonads. Nat Biotechnol 4, 439–444 (1986) doi:10.1038/nbt0586-439

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