Citation:

Walter, M., L. Porteous, AND R. Seidler. EVALUATION OF A METHOD TO MEASURE CONJUGAL TRANSFER OF RECOMBINANT DNA IN SOIL SLURRIES. U.S. Environmental Protection Agency, Washington, D.C., EPA/600/J-89/480 (NTIS PB91117580), 1989.

Description:

Release of recombinant microbes into the environment necessitates an evaluation of their ability to transfer genetic material. he present report evaluates a method to detect conjugal DNA plasmid transfer in soil slurries under various environmental conditions. onor Pseudomonas cepacia containing pR388::Tn1721 end P. cepacia recipient cultures were co-incubated in soil slurries containing autoclaved or natural soil end treated with one or more of 14 experimental conditions. onjugal mating frequency (transconjugants per initial donor) ranged from 4.8 x 10- 1 to 1.9 x 10- 7. ighest numbers of transconjugants, 1.5 x 10 7 colony forming units/ml soil slurry, were observed following incubation t 35 degrees C with an enriched nutrient supplement added to the soil. ow numbers of transconjugants, 10 3 colony forming units/ml soil slurry, were observed when mating pairs were subjected to low nutrient or pH stress even though initial donor end recipient populations were maintained at high levels. his test system provides a simple way to estimate effects of changing environmental factors on plasmid transfer rates end on the survival of recombinant microorganisms. y use of soil collected from sites proposed to receive genetically engineered microorganisms, preliminary risk assessments can be obtained regarding the potential for gene transfer and microorganism survival with this soil slurry test system.

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