Effectiveness of Chromosomal and Plasmid-linked Genes for Enumerating Biotechnology Agents In Vivo

Article English OPEN
George, S. Elizabeth ; Nelson, Gail M. (2011)
  • Publisher: Microbial Ecology in Health and Disease
  • Journal: Microbial Ecology in Health and Disease (issn: 1651-2235)
  • Related identifiers: doi: 10.3402/mehd.v14i1.8217

Following the release of biotechnology agents, detection methods for monitoring human exposure to these microorganisms should be available. Enzymatic markers from chromosomal (lacZY) and plasmid (xylE) DNA were evaluated in an in vivo mouse model. Mice were treated orally or intranasally with Pseudomonas aureofaciens 3732RN-L11 (lacZY), 3732RN-L11::pRO1940 (lacZY xylE), or ATCC 13985. All strains were cleared within 5 days from the cecum or lungs. Direct plating, without intermediate antibiotic resistance selection, was used to detect the genetic markers. Chromosomal-linked lacZ was stable in vivo and in vitro but plasmid-linked xylE was not. Of the isolates recovered, 75% (in vivo, oral), 98% (in vivo, intranasal), and 16% (in vitro) remained XyIE+. Though more tedious to enumerate, both lacZY and xylE are plausible alternatives to antibiotic resistance markers for in vivo biotechnology risk assessment studies.Keywords: Biotechnology Risk Assessment, Pseudomonas aureofaciens, Colonization in vivo.
  • References (30)
    30 references, page 1 of 3

    1. Levy SB, Marshall B, Rowse-Eagle D. Survival of Escherichia coli host-vector systems in the mammalian intestine. Science 1980; 209: 391 - 4.

    2. Lindow SE, Knudsen GR, Seidler RJ, Walter MV, Lambou VW, Amy PS, Schmedding D, Prince V, Hern S. Aerial dispersal and epiphytic survival of Pseudomonas syringae during a pretest for the release of genetically engineered strains into the environment. Appl Environ Microbiol 1988; 54: 1557 - 63.

    3. Cohen SP, Yan W, Levy SB. A multidrug resistance regulatory chromosomal locus is widespread among enteric bacteria. J Infect Dis 1993; 168: 484 - 8.

    4. Marshall B, Petrowski D, Levy SB. Inter- and intraspecies spread of Escherichia coli in a farm environment in the absence of antibiotic usage. Proc Natl Acad Sci USA 1990; 87: 6609 - 13.

    5. Federal Register. Microbial Products of Biotechnology; proposed regulation under the toxic substances control act; proposed rule. Federal Register 1994; 59: 45526 - 85.

    6. Bolivar F, Rodriques RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW, Crosa JH, Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 1977; 2: 95- 113.

    7. Curtiss R, III, Inoue M, Pereira D, Hsu JC, Alexander L, Rock L. Construction and use of safer bacteria host strains for recombinant DNA research. In: Scott WA, Werner R, eds. Molecular Cloning of Recombinant DNA, Miami Winter Symposia, volume 13. New York: Academic Press, 1977: 99 - 114.

    8. Saylers AA, Shoemaker NB, Stevens AM, Li L-Y. Conjugative transposons: an unusual and diverse set of integrated gene transfer elements. Microbiol Revs 1995; 59: 579 - 90.

    9. Shoemaker NB, Vlamakis H, Hayes K, Salyers AA. Evidence for extensive resistance gene transfer among Bacteriodes spp. and among Bacteriodes and other genera in the human colon. Appl Environ Microbiol 2001; 67: 561 - 8.

    10. Haack BJ, Andrews RE, Loynachan TE. Conjugal transfer on Tn916 in model soil systems. In: Levin M, Grim C, Angle JS, eds. Biotechnology Risk Assessment: USEPA:USDA:Environment Canada, Proceedings of the Biotechnology Risk Assessment Symposium. College Park, MD: University of Maryland, 1994: 45- 9.

  • Metrics
    No metrics available
Share - Bookmark