Detection and Enumeration of a Tagged Pseudomonas fluorescens Strain by Using Soil with Markers Associated with an Engineered Catabolic Pathway.

Applied and Environmental Microbiology
I Hwang, S K Farrand

Abstract

Previously we described a novel gene tagging method, using the moc (mannityl opine catabolism) region from the Agrobacterium tumefaciens Ti plasmid pTi15955, to identify microorganisms destined for release into the environment. Here, we used the engineered strain Pseudomonas fluorescens PF5MT12 carrying the moc region integrated into the bacterial chromosome to demonstrate the usefulness of the markers for detection and direct selection of marked organisms present in soil samples. Using this system, we routinely detected population levels as low as 10(sup2) CFU per g of soil sampled. In addition to direct selection, we developed an immunologically based assay using MOP cyclase, a unique enzyme associated with moc, as the epitope for detecting the tagged organism. The colony immunoblot assay proved to be highly specific and without any false-positive signals when used to identify organisms cultured from soil on nonselective medium. The numbers of colonies that were immunoreactive with the anti-MOP cyclase antibody were essentially equal to those that grew out on selection plates. This indicates that MOP cyclase can be used as a marker and that we can use nonselective medium to retrieve the marked genetically engineered microorga...Continue Reading

References

Jan 1, 1992·Applied and Environmental Microbiology·J J ShawJ W Kloepper
Feb 1, 1992·PCR Methods and Applications·A K Bej, M H Mahbubani
Feb 1, 1992·Applied and Environmental Microbiology·Y L Tsai, B H Olson
Jan 1, 1991·Annual Review of Microbiology·R J Steffan, R M Atlas
Dec 1, 1991·Applied and Environmental Microbiology·A K BejR M Atlas
Feb 1, 1990·Applied and Environmental Microbiology·A K BejR M Atlas
May 1, 1989·Applied and Environmental Microbiology·G R ChaudhryA R Bhatti
Jun 1, 1987·Molecular & General Genetics : MGG·Y DessauxS K Farrand
Sep 1, 1988·Applied and Environmental Microbiology·R J Steffan, R M Atlas
Mar 1, 1994·Applied and Environmental Microbiology·I Hwang, S K Farrand
Feb 1, 1989·Applied and Environmental Microbiology·P E Olsen, W A Rice
Aug 1, 1990·Applied and Environmental Microbiology·C S Nautiyal, P Dion

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Citations

Nov 17, 2010·Journal of Agricultural and Food Chemistry·Sung Eun KimSung Uk Kim

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